Www.tandfonline.com



Supplementary InformationA general strategy for generating intact, full-length IgG antibodies that penetrate into the cytosol of living cellsDong-Ki Choi, Jeomil Bae, Seung-Min Shin, Ju-Yeon Shin, Sunghoon Kim, and Yong-Sung KimInventory of Supplementary InformationSupplementary Figure legends S1 to S8Supplementary Movie S1 Supplementary Materials and MethodsSupplementary ReferencesSupplementary Figure S1. (A) Amino acid sequence alignment of m3D8, hT0, hT2 and hT3 VL single domain antibodies. CDR residues are indicated by brackets ([ ]). The Vernier zone residues in the β-sheet framework closely underlying CDRs are underlined and bolded. The two Vernier zone residues back mutated in hT2 and hT3 VLs with the corresponding murine residues (I2L and L4M) are indicated by bold, red color. Amino acids changes in Vκ1 framework of hT3 VL from Vκ3 framework of hT2 VL are colored by blue. Residues are numbered according to the Kabat numbering system. ADDIN EN.CITE <EndNote><Cite><Author>Kabat</Author><Year>1991</Year><RecNum>403</RecNum><DisplayText><style face="superscript">1</style></DisplayText><record><rec-number>403</rec-number><foreign-keys><key app="EN" db-id="5sz0e52zssawxcea2xppw558frz5zzs2d02v">403</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Kabat, E. A.</author><author>Wu, T. T.</author></authors></contributors><auth-address>Department of Microbiology, College of Physicians and Surgeons, Columbia University, New York, NY 10032.</auth-address><titles><title>Identical V region amino acid sequences and segments of sequences in antibodies of different specificities. Relative contributions of VH and VL genes, minigenes, and complementarity-determining regions to binding of antibody-combining sites</title><secondary-title>J Immunol</secondary-title><alt-title>Journal of immunology</alt-title></titles><periodical><full-title>J Immunol</full-title></periodical><pages>1709-19</pages><volume>147</volume><number>5</number><keywords><keyword>Amino Acid Sequence</keyword><keyword>Animals</keyword><keyword>*Antibody Specificity</keyword><keyword>*Binding Sites, Antibody</keyword><keyword>*Genes, Immunoglobulin</keyword><keyword>Guinea Pigs</keyword><keyword>Humans</keyword><keyword>Immunoglobulin Heavy Chains/*genetics</keyword><keyword>Immunoglobulin Light Chains/*genetics</keyword><keyword>Immunoglobulin Variable Region/*chemistry/genetics/immunology</keyword><keyword>Mice</keyword><keyword>Molecular Sequence Data</keyword><keyword>Protein Conformation</keyword><keyword>Rabbits</keyword><keyword>Rats</keyword></keywords><dates><year>1991</year><pub-dates><date>Sep 1</date></pub-dates></dates><isbn>0022-1767 (Print)&#xD;0022-1767 (Linking)</isbn><accession-num>1908882</accession-num><urls><related-urls><url> (B and C) The humanized hT2 and hT3 VLs retained the caveolae-mediated endocytosis pathway of m3D8 VL, determined by co-localization of internalized VLs with endocytosis markers (B) and effects of endocytosis inhibitors on the cellular uptake (C). In (B), HeLa cells were co-treated at 37 °C for 2 h with VLs (10 M) and endocytosis markers including 10 g/ml Alexa Fluor 488-transferrin (TF) for clathrin-mediated endocytosis, FITC-cholera toxin B (Ctx-B) for caveolae-mediated endocytosis, and Oregon green-dextran (Dextran) for macropinocytosis PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5KYW5nPC9BdXRob3I+PFllYXI+MjAwOTwvWWVhcj48UmVj

TnVtPjIwPC9SZWNOdW0+PERpc3BsYXlUZXh0PjxzdHlsZSBmYWNlPSJzdXBlcnNjcmlwdCI+Miwg

Mzwvc3R5bGU+PC9EaXNwbGF5VGV4dD48cmVjb3JkPjxyZWMtbnVtYmVyPjIwPC9yZWMtbnVtYmVy

Pjxmb3JlaWduLWtleXM+PGtleSBhcHA9IkVOIiBkYi1pZD0icnpzZTB3c2Y5cnd4dzhldnphbnBh

dmY5YXp6OTJkd2RhdHRzIj4yMDwva2V5PjwvZm9yZWlnbi1rZXlzPjxyZWYtdHlwZSBuYW1lPSJK

b3VybmFsIEFydGljbGUiPjE3PC9yZWYtdHlwZT48Y29udHJpYnV0b3JzPjxhdXRob3JzPjxhdXRo

b3I+SmFuZywgSi4gWS48L2F1dGhvcj48YXV0aG9yPkplb25nLCBKLiBHLjwvYXV0aG9yPjxhdXRo

b3I+SnVuLCBILiBSLjwvYXV0aG9yPjxhdXRob3I+TGVlLCBTLiBDLjwvYXV0aG9yPjxhdXRob3I+

S2ltLCBKLiBTLjwvYXV0aG9yPjxhdXRob3I+S2ltLCBZLiBTLjwvYXV0aG9yPjxhdXRob3I+S3dv

biwgTS4gSC48L2F1dGhvcj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVzcz5E

ZXBhcnRtZW50IG9mIE1pY3JvYmlvbG9neSwgQWpvdSBVbml2ZXJzaXR5IFNjaG9vbCBvZiBNZWRp

Y2luZSwgU3V3b24sIDQ0My03MjEsIEtvcmVhLjwvYXV0aC1hZGRyZXNzPjx0aXRsZXM+PHRpdGxl

PkEgbnVjbGVpYyBhY2lkLWh5ZHJvbHl6aW5nIGFudGlib2R5IHBlbmV0cmF0ZXMgaW50byBjZWxs

cyB2aWEgY2F2ZW9sYWUtbWVkaWF0ZWQgZW5kb2N5dG9zaXMsIGxvY2FsaXplcyBpbiB0aGUgY3l0

b3NvbCBhbmQgZXhoaWJpdHMgY3l0b3RveGljaXR5PC90aXRsZT48c2Vjb25kYXJ5LXRpdGxlPkNl

bGwgTW9sIExpZmUgU2NpPC9zZWNvbmRhcnktdGl0bGU+PGFsdC10aXRsZT5DZWxsdWxhciBhbmQg

bW9sZWN1bGFyIGxpZmUgc2NpZW5jZXMgOiBDTUxTPC9hbHQtdGl0bGU+PC90aXRsZXM+PHBlcmlv

ZGljYWw+PGZ1bGwtdGl0bGU+Q2VsbCBNb2wgTGlmZSBTY2k8L2Z1bGwtdGl0bGU+PGFiYnItMT5D

ZWxsdWxhciBhbmQgbW9sZWN1bGFyIGxpZmUgc2NpZW5jZXMgOiBDTUxTPC9hYmJyLTE+PC9wZXJp

b2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVsbC10aXRsZT5DZWxsIE1vbCBMaWZlIFNjaTwvZnVs

bC10aXRsZT48YWJici0xPkNlbGx1bGFyIGFuZCBtb2xlY3VsYXIgbGlmZSBzY2llbmNlcyA6IENN

TFM8L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4xOTg1LTk3PC9wYWdlcz48dm9sdW1l

PjY2PC92b2x1bWU+PG51bWJlcj4xMS0xMjwvbnVtYmVyPjxrZXl3b3Jkcz48a2V5d29yZD5BbnRp

Ym9kaWVzLCBBbnRpbnVjbGVhci8qbWV0YWJvbGlzbS9waGFybWFjb2xvZ3k8L2tleXdvcmQ+PGtl

eXdvcmQ+QXBvcHRvc2lzPC9rZXl3b3JkPjxrZXl3b3JkPkNhdmVvbGFlLypwaHlzaW9sb2d5PC9r

ZXl3b3JkPjxrZXl3b3JkPkNlbGwgTWVtYnJhbmUgUGVybWVhYmlsaXR5PC9rZXl3b3JkPjxrZXl3

b3JkPkNlbGwgU3Vydml2YWwvZHJ1ZyBlZmZlY3RzPC9rZXl3b3JkPjxrZXl3b3JkPkN5dG9zb2wv

Km1ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+RW5kb2N5dG9zaXM8L2tleXdvcmQ+PGtleXdv

cmQ+SGVMYSBDZWxsczwva2V5d29yZD48a2V5d29yZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdvcmQ+

SHlkcm9seXNpczwva2V5d29yZD48a2V5d29yZD5JbW11bm9nbG9idWxpbiBWYXJpYWJsZSBSZWdp

b24vKm1ldGFib2xpc20vcGhhcm1hY29sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPk1lbWJyYW5lIE1p

Y3JvZG9tYWluczwva2V5d29yZD48a2V5d29yZD5Qcm90ZW9nbHljYW5zL21ldGFib2xpc208L2tl

eXdvcmQ+PGtleXdvcmQ+Uk5BL21ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+Umlib251Y2xl

YXNlcy9tZXRhYm9saXNtPC9rZXl3b3JkPjwva2V5d29yZHM+PGRhdGVzPjx5ZWFyPjIwMDk8L3ll

YXI+PHB1Yi1kYXRlcz48ZGF0ZT5KdW48L2RhdGU+PC9wdWItZGF0ZXM+PC9kYXRlcz48aXNibj4x

NDIwLTkwNzEgKEVsZWN0cm9uaWMpJiN4RDsxNDIwLTY4MlggKExpbmtpbmcpPC9pc2JuPjxhY2Nl

c3Npb24tbnVtPjE5MzczNDM0PC9hY2Nlc3Npb24tbnVtPjx1cmxzPjxyZWxhdGVkLXVybHM+PHVy

bD5odHRwOi8vd3d3Lm5jYmkubmxtLm5paC5nb3YvcHVibWVkLzE5MzczNDM0PC91cmw+PC9yZWxh

dGVkLXVybHM+PC91cmxzPjxlbGVjdHJvbmljLXJlc291cmNlLW51bT4xMC4xMDA3L3MwMDAxOC0w

MDktOTE3OS0yPC9lbGVjdHJvbmljLXJlc291cmNlLW51bT48L3JlY29yZD48L0NpdGU+PENpdGU+

PEF1dGhvcj5MZWU8L0F1dGhvcj48WWVhcj4yMDEwPC9ZZWFyPjxSZWNOdW0+MjI8L1JlY051bT48

cmVjb3JkPjxyZWMtbnVtYmVyPjIyPC9yZWMtbnVtYmVyPjxmb3JlaWduLWtleXM+PGtleSBhcHA9

IkVOIiBkYi1pZD0iNXN6MGU1Mnpzc2F3eGNlYTJ4cHB3NTU4ZnJ6NXp6czJkMDJ2Ij4yMjwva2V5

PjwvZm9yZWlnbi1rZXlzPjxyZWYtdHlwZSBuYW1lPSJKb3VybmFsIEFydGljbGUiPjE3PC9yZWYt

dHlwZT48Y29udHJpYnV0b3JzPjxhdXRob3JzPjxhdXRob3I+TGVlLCBXLiBSLjwvYXV0aG9yPjxh

dXRob3I+SmFuZywgSi4gWS48L2F1dGhvcj48YXV0aG9yPktpbSwgSi4gUy48L2F1dGhvcj48YXV0

aG9yPkt3b24sIE0uIEguPC9hdXRob3I+PGF1dGhvcj5LaW0sIFkuIFMuPC9hdXRob3I+PC9hdXRo

b3JzPjwvY29udHJpYnV0b3JzPjxhdXRoLWFkZHJlc3M+RGVwYXJ0bWVudCBvZiBNb2xlY3VsYXIg

U2NpZW5jZSBhbmQgVGVjaG5vbG9neSwgQWpvdSBVbml2ZXJzaXR5LCBTdXdvbiA0NDMtNzQ5LCBL

b3JlYS48L2F1dGgtYWRkcmVzcz48dGl0bGVzPjx0aXRsZT5HZW5lIHNpbGVuY2luZyBieSBjZWxs

LXBlbmV0cmF0aW5nLCBzZXF1ZW5jZS1zZWxlY3RpdmUgYW5kIG51Y2xlaWMtYWNpZCBoeWRyb2x5

emluZyBhbnRpYm9kaWVzPC90aXRsZT48c2Vjb25kYXJ5LXRpdGxlPk51Y2xlaWMgQWNpZHMgUmVz

PC9zZWNvbmRhcnktdGl0bGU+PGFsdC10aXRsZT5OdWNsZWljIGFjaWRzIHJlc2VhcmNoPC9hbHQt

dGl0bGU+PC90aXRsZXM+PHBlcmlvZGljYWw+PGZ1bGwtdGl0bGU+TnVjbGVpYyBBY2lkcyBSZXM8

L2Z1bGwtdGl0bGU+PGFiYnItMT5OdWNsZWljIGFjaWRzIHJlc2VhcmNoPC9hYmJyLTE+PC9wZXJp

b2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVsbC10aXRsZT5OdWNsZWljIEFjaWRzIFJlczwvZnVs

bC10aXRsZT48YWJici0xPk51Y2xlaWMgYWNpZHMgcmVzZWFyY2g8L2FiYnItMT48L2FsdC1wZXJp

b2RpY2FsPjxwYWdlcz4xNTk2LTYwOTwvcGFnZXM+PHZvbHVtZT4zODwvdm9sdW1lPjxudW1iZXI+

NTwvbnVtYmVyPjxlZGl0aW9uPjIwMDkvMTIvMTc8L2VkaXRpb24+PGtleXdvcmRzPjxrZXl3b3Jk

PkFudGlib2RpZXMsIENhdGFseXRpYy9jaGVtaXN0cnkvKm1ldGFib2xpc208L2tleXdvcmQ+PGtl

eXdvcmQ+QXBvcHRvc2lzPC9rZXl3b3JkPjxrZXl3b3JkPkJhc2UgU2VxdWVuY2U8L2tleXdvcmQ+

PGtleXdvcmQ+Q2VsbCBMaW5lLCBUdW1vcjwva2V5d29yZD48a2V5d29yZD5DeXRvc29sL21ldGFi

b2xpc208L2tleXdvcmQ+PGtleXdvcmQ+RE5BLCBTaW5nbGUtU3RyYW5kZWQvbWV0YWJvbGlzbTwv

a2V5d29yZD48a2V5d29yZD5HZW5lIEtub2NrZG93biBUZWNobmlxdWVzPC9rZXl3b3JkPjxrZXl3

b3JkPkdlbmUgTGlicmFyeTwva2V5d29yZD48a2V5d29yZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdv

cmQ+SHlkcm9seXNpczwva2V5d29yZD48a2V5d29yZD5JbW11bm9nbG9idWxpbiBMaWdodCBDaGFp

bnMvY2hlbWlzdHJ5PC9rZXl3b3JkPjxrZXl3b3JkPkltbXVub2dsb2J1bGluIFZhcmlhYmxlIFJl

Z2lvbi9jaGVtaXN0cnk8L2tleXdvcmQ+PGtleXdvcmQ+KlJOQSBJbnRlcmZlcmVuY2U8L2tleXdv

cmQ+PGtleXdvcmQ+Uk5BLCBNZXNzZW5nZXIvKm1ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+

UmVjZXB0b3IsIGVyYkItMi9hbnRhZ29uaXN0cyAmYW1wOyBpbmhpYml0b3JzL2dlbmV0aWNzL21l

dGFib2xpc208L2tleXdvcmQ+PC9rZXl3b3Jkcz48ZGF0ZXM+PHllYXI+MjAxMDwveWVhcj48cHVi

LWRhdGVzPjxkYXRlPk1hcjwvZGF0ZT48L3B1Yi1kYXRlcz48L2RhdGVzPjxpc2JuPjEzNjItNDk2

MiAoRWxlY3Ryb25pYykmI3hEOzAzMDUtMTA0OCAoTGlua2luZyk8L2lzYm4+PGFjY2Vzc2lvbi1u

dW0+MjAwMDc2MDI8L2FjY2Vzc2lvbi1udW0+PHdvcmstdHlwZT5SZXNlYXJjaCBTdXBwb3J0LCBO

b24tVS5TLiBHb3YmYXBvczt0PC93b3JrLXR5cGU+PHVybHM+PHJlbGF0ZWQtdXJscz48dXJsPmh0

dHA6Ly93d3cubmNiaS5ubG0ubmloLmdvdi9wdWJtZWQvMjAwMDc2MDI8L3VybD48L3JlbGF0ZWQt

dXJscz48L3VybHM+PGN1c3RvbTI+MjgzNjU3MjwvY3VzdG9tMj48ZWxlY3Ryb25pYy1yZXNvdXJj

ZS1udW0+MTAuMTA5My9uYXIvZ2twMTE0NTwvZWxlY3Ryb25pYy1yZXNvdXJjZS1udW0+PGxhbmd1

YWdlPmVuZzwvbGFuZ3VhZ2U+PC9yZWNvcmQ+PC9DaXRlPjwvRW5kTm90ZT5=

ADDIN EN.CITE PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5KYW5nPC9BdXRob3I+PFllYXI+MjAwOTwvWWVhcj48UmVj

TnVtPjIwPC9SZWNOdW0+PERpc3BsYXlUZXh0PjxzdHlsZSBmYWNlPSJzdXBlcnNjcmlwdCI+Miwg

Mzwvc3R5bGU+PC9EaXNwbGF5VGV4dD48cmVjb3JkPjxyZWMtbnVtYmVyPjIwPC9yZWMtbnVtYmVy

Pjxmb3JlaWduLWtleXM+PGtleSBhcHA9IkVOIiBkYi1pZD0icnpzZTB3c2Y5cnd4dzhldnphbnBh

dmY5YXp6OTJkd2RhdHRzIj4yMDwva2V5PjwvZm9yZWlnbi1rZXlzPjxyZWYtdHlwZSBuYW1lPSJK

b3VybmFsIEFydGljbGUiPjE3PC9yZWYtdHlwZT48Y29udHJpYnV0b3JzPjxhdXRob3JzPjxhdXRo

b3I+SmFuZywgSi4gWS48L2F1dGhvcj48YXV0aG9yPkplb25nLCBKLiBHLjwvYXV0aG9yPjxhdXRo

b3I+SnVuLCBILiBSLjwvYXV0aG9yPjxhdXRob3I+TGVlLCBTLiBDLjwvYXV0aG9yPjxhdXRob3I+

S2ltLCBKLiBTLjwvYXV0aG9yPjxhdXRob3I+S2ltLCBZLiBTLjwvYXV0aG9yPjxhdXRob3I+S3dv

biwgTS4gSC48L2F1dGhvcj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVzcz5E

ZXBhcnRtZW50IG9mIE1pY3JvYmlvbG9neSwgQWpvdSBVbml2ZXJzaXR5IFNjaG9vbCBvZiBNZWRp

Y2luZSwgU3V3b24sIDQ0My03MjEsIEtvcmVhLjwvYXV0aC1hZGRyZXNzPjx0aXRsZXM+PHRpdGxl

PkEgbnVjbGVpYyBhY2lkLWh5ZHJvbHl6aW5nIGFudGlib2R5IHBlbmV0cmF0ZXMgaW50byBjZWxs

cyB2aWEgY2F2ZW9sYWUtbWVkaWF0ZWQgZW5kb2N5dG9zaXMsIGxvY2FsaXplcyBpbiB0aGUgY3l0

b3NvbCBhbmQgZXhoaWJpdHMgY3l0b3RveGljaXR5PC90aXRsZT48c2Vjb25kYXJ5LXRpdGxlPkNl

bGwgTW9sIExpZmUgU2NpPC9zZWNvbmRhcnktdGl0bGU+PGFsdC10aXRsZT5DZWxsdWxhciBhbmQg

bW9sZWN1bGFyIGxpZmUgc2NpZW5jZXMgOiBDTUxTPC9hbHQtdGl0bGU+PC90aXRsZXM+PHBlcmlv

ZGljYWw+PGZ1bGwtdGl0bGU+Q2VsbCBNb2wgTGlmZSBTY2k8L2Z1bGwtdGl0bGU+PGFiYnItMT5D

ZWxsdWxhciBhbmQgbW9sZWN1bGFyIGxpZmUgc2NpZW5jZXMgOiBDTUxTPC9hYmJyLTE+PC9wZXJp

b2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVsbC10aXRsZT5DZWxsIE1vbCBMaWZlIFNjaTwvZnVs

bC10aXRsZT48YWJici0xPkNlbGx1bGFyIGFuZCBtb2xlY3VsYXIgbGlmZSBzY2llbmNlcyA6IENN

TFM8L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4xOTg1LTk3PC9wYWdlcz48dm9sdW1l

PjY2PC92b2x1bWU+PG51bWJlcj4xMS0xMjwvbnVtYmVyPjxrZXl3b3Jkcz48a2V5d29yZD5BbnRp

Ym9kaWVzLCBBbnRpbnVjbGVhci8qbWV0YWJvbGlzbS9waGFybWFjb2xvZ3k8L2tleXdvcmQ+PGtl

eXdvcmQ+QXBvcHRvc2lzPC9rZXl3b3JkPjxrZXl3b3JkPkNhdmVvbGFlLypwaHlzaW9sb2d5PC9r

ZXl3b3JkPjxrZXl3b3JkPkNlbGwgTWVtYnJhbmUgUGVybWVhYmlsaXR5PC9rZXl3b3JkPjxrZXl3

b3JkPkNlbGwgU3Vydml2YWwvZHJ1ZyBlZmZlY3RzPC9rZXl3b3JkPjxrZXl3b3JkPkN5dG9zb2wv

Km1ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+RW5kb2N5dG9zaXM8L2tleXdvcmQ+PGtleXdv

cmQ+SGVMYSBDZWxsczwva2V5d29yZD48a2V5d29yZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdvcmQ+

SHlkcm9seXNpczwva2V5d29yZD48a2V5d29yZD5JbW11bm9nbG9idWxpbiBWYXJpYWJsZSBSZWdp

b24vKm1ldGFib2xpc20vcGhhcm1hY29sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPk1lbWJyYW5lIE1p

Y3JvZG9tYWluczwva2V5d29yZD48a2V5d29yZD5Qcm90ZW9nbHljYW5zL21ldGFib2xpc208L2tl

eXdvcmQ+PGtleXdvcmQ+Uk5BL21ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+Umlib251Y2xl

YXNlcy9tZXRhYm9saXNtPC9rZXl3b3JkPjwva2V5d29yZHM+PGRhdGVzPjx5ZWFyPjIwMDk8L3ll

YXI+PHB1Yi1kYXRlcz48ZGF0ZT5KdW48L2RhdGU+PC9wdWItZGF0ZXM+PC9kYXRlcz48aXNibj4x

NDIwLTkwNzEgKEVsZWN0cm9uaWMpJiN4RDsxNDIwLTY4MlggKExpbmtpbmcpPC9pc2JuPjxhY2Nl

c3Npb24tbnVtPjE5MzczNDM0PC9hY2Nlc3Npb24tbnVtPjx1cmxzPjxyZWxhdGVkLXVybHM+PHVy

bD5odHRwOi8vd3d3Lm5jYmkubmxtLm5paC5nb3YvcHVibWVkLzE5MzczNDM0PC91cmw+PC9yZWxh

dGVkLXVybHM+PC91cmxzPjxlbGVjdHJvbmljLXJlc291cmNlLW51bT4xMC4xMDA3L3MwMDAxOC0w

MDktOTE3OS0yPC9lbGVjdHJvbmljLXJlc291cmNlLW51bT48L3JlY29yZD48L0NpdGU+PENpdGU+

PEF1dGhvcj5MZWU8L0F1dGhvcj48WWVhcj4yMDEwPC9ZZWFyPjxSZWNOdW0+MjI8L1JlY051bT48

cmVjb3JkPjxyZWMtbnVtYmVyPjIyPC9yZWMtbnVtYmVyPjxmb3JlaWduLWtleXM+PGtleSBhcHA9

IkVOIiBkYi1pZD0iNXN6MGU1Mnpzc2F3eGNlYTJ4cHB3NTU4ZnJ6NXp6czJkMDJ2Ij4yMjwva2V5

PjwvZm9yZWlnbi1rZXlzPjxyZWYtdHlwZSBuYW1lPSJKb3VybmFsIEFydGljbGUiPjE3PC9yZWYt

dHlwZT48Y29udHJpYnV0b3JzPjxhdXRob3JzPjxhdXRob3I+TGVlLCBXLiBSLjwvYXV0aG9yPjxh

dXRob3I+SmFuZywgSi4gWS48L2F1dGhvcj48YXV0aG9yPktpbSwgSi4gUy48L2F1dGhvcj48YXV0

aG9yPkt3b24sIE0uIEguPC9hdXRob3I+PGF1dGhvcj5LaW0sIFkuIFMuPC9hdXRob3I+PC9hdXRo

b3JzPjwvY29udHJpYnV0b3JzPjxhdXRoLWFkZHJlc3M+RGVwYXJ0bWVudCBvZiBNb2xlY3VsYXIg

U2NpZW5jZSBhbmQgVGVjaG5vbG9neSwgQWpvdSBVbml2ZXJzaXR5LCBTdXdvbiA0NDMtNzQ5LCBL

b3JlYS48L2F1dGgtYWRkcmVzcz48dGl0bGVzPjx0aXRsZT5HZW5lIHNpbGVuY2luZyBieSBjZWxs

LXBlbmV0cmF0aW5nLCBzZXF1ZW5jZS1zZWxlY3RpdmUgYW5kIG51Y2xlaWMtYWNpZCBoeWRyb2x5

emluZyBhbnRpYm9kaWVzPC90aXRsZT48c2Vjb25kYXJ5LXRpdGxlPk51Y2xlaWMgQWNpZHMgUmVz

PC9zZWNvbmRhcnktdGl0bGU+PGFsdC10aXRsZT5OdWNsZWljIGFjaWRzIHJlc2VhcmNoPC9hbHQt

dGl0bGU+PC90aXRsZXM+PHBlcmlvZGljYWw+PGZ1bGwtdGl0bGU+TnVjbGVpYyBBY2lkcyBSZXM8

L2Z1bGwtdGl0bGU+PGFiYnItMT5OdWNsZWljIGFjaWRzIHJlc2VhcmNoPC9hYmJyLTE+PC9wZXJp

b2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVsbC10aXRsZT5OdWNsZWljIEFjaWRzIFJlczwvZnVs

bC10aXRsZT48YWJici0xPk51Y2xlaWMgYWNpZHMgcmVzZWFyY2g8L2FiYnItMT48L2FsdC1wZXJp

b2RpY2FsPjxwYWdlcz4xNTk2LTYwOTwvcGFnZXM+PHZvbHVtZT4zODwvdm9sdW1lPjxudW1iZXI+

NTwvbnVtYmVyPjxlZGl0aW9uPjIwMDkvMTIvMTc8L2VkaXRpb24+PGtleXdvcmRzPjxrZXl3b3Jk

PkFudGlib2RpZXMsIENhdGFseXRpYy9jaGVtaXN0cnkvKm1ldGFib2xpc208L2tleXdvcmQ+PGtl

eXdvcmQ+QXBvcHRvc2lzPC9rZXl3b3JkPjxrZXl3b3JkPkJhc2UgU2VxdWVuY2U8L2tleXdvcmQ+

PGtleXdvcmQ+Q2VsbCBMaW5lLCBUdW1vcjwva2V5d29yZD48a2V5d29yZD5DeXRvc29sL21ldGFi

b2xpc208L2tleXdvcmQ+PGtleXdvcmQ+RE5BLCBTaW5nbGUtU3RyYW5kZWQvbWV0YWJvbGlzbTwv

a2V5d29yZD48a2V5d29yZD5HZW5lIEtub2NrZG93biBUZWNobmlxdWVzPC9rZXl3b3JkPjxrZXl3

b3JkPkdlbmUgTGlicmFyeTwva2V5d29yZD48a2V5d29yZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdv

cmQ+SHlkcm9seXNpczwva2V5d29yZD48a2V5d29yZD5JbW11bm9nbG9idWxpbiBMaWdodCBDaGFp

bnMvY2hlbWlzdHJ5PC9rZXl3b3JkPjxrZXl3b3JkPkltbXVub2dsb2J1bGluIFZhcmlhYmxlIFJl

Z2lvbi9jaGVtaXN0cnk8L2tleXdvcmQ+PGtleXdvcmQ+KlJOQSBJbnRlcmZlcmVuY2U8L2tleXdv

cmQ+PGtleXdvcmQ+Uk5BLCBNZXNzZW5nZXIvKm1ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+

UmVjZXB0b3IsIGVyYkItMi9hbnRhZ29uaXN0cyAmYW1wOyBpbmhpYml0b3JzL2dlbmV0aWNzL21l

dGFib2xpc208L2tleXdvcmQ+PC9rZXl3b3Jkcz48ZGF0ZXM+PHllYXI+MjAxMDwveWVhcj48cHVi

LWRhdGVzPjxkYXRlPk1hcjwvZGF0ZT48L3B1Yi1kYXRlcz48L2RhdGVzPjxpc2JuPjEzNjItNDk2

MiAoRWxlY3Ryb25pYykmI3hEOzAzMDUtMTA0OCAoTGlua2luZyk8L2lzYm4+PGFjY2Vzc2lvbi1u

dW0+MjAwMDc2MDI8L2FjY2Vzc2lvbi1udW0+PHdvcmstdHlwZT5SZXNlYXJjaCBTdXBwb3J0LCBO

b24tVS5TLiBHb3YmYXBvczt0PC93b3JrLXR5cGU+PHVybHM+PHJlbGF0ZWQtdXJscz48dXJsPmh0

dHA6Ly93d3cubmNiaS5ubG0ubmloLmdvdi9wdWJtZWQvMjAwMDc2MDI8L3VybD48L3JlbGF0ZWQt

dXJscz48L3VybHM+PGN1c3RvbTI+MjgzNjU3MjwvY3VzdG9tMj48ZWxlY3Ryb25pYy1yZXNvdXJj

ZS1udW0+MTAuMTA5My9uYXIvZ2twMTE0NTwvZWxlY3Ryb25pYy1yZXNvdXJjZS1udW0+PGxhbmd1

YWdlPmVuZzwvbGFuZ3VhZ2U+PC9yZWNvcmQ+PC9DaXRlPjwvRW5kTm90ZT5=

ADDIN EN.CITE.DATA 2, 3 prior to the confocal microscopy analysis. In (C), HeLa cells were pre-treated for 30 min with CPZ (10 μg/ml), MβCD (5 mM) or Cyt-D (1 μg/ml) and then incubated each VL for additional for 2 h, followed by confocal microscopy analysis. In (B and C), internalized Protein-A-tagged VLs were detected with rabbit IgG and subsequent TRITC-conjugated goat anti-rabbit antibody.PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5MZWU8L0F1dGhvcj48WWVhcj4yMDEwPC9ZZWFyPjxSZWNO

dW0+MjI8L1JlY051bT48RGlzcGxheVRleHQ+PHN0eWxlIGZhY2U9InN1cGVyc2NyaXB0Ij4zPC9z

dHlsZT48L0Rpc3BsYXlUZXh0PjxyZWNvcmQ+PHJlYy1udW1iZXI+MjI8L3JlYy1udW1iZXI+PGZv

cmVpZ24ta2V5cz48a2V5IGFwcD0iRU4iIGRiLWlkPSI1c3owZTUyenNzYXd4Y2VhMnhwcHc1NThm

cno1enpzMmQwMnYiPjIyPC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10eXBlIG5hbWU9IkpvdXJu

YWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1dGhvcnM+PGF1dGhvcj5M

ZWUsIFcuIFIuPC9hdXRob3I+PGF1dGhvcj5KYW5nLCBKLiBZLjwvYXV0aG9yPjxhdXRob3I+S2lt

LCBKLiBTLjwvYXV0aG9yPjxhdXRob3I+S3dvbiwgTS4gSC48L2F1dGhvcj48YXV0aG9yPktpbSwg

WS4gUy48L2F1dGhvcj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVzcz5EZXBh

cnRtZW50IG9mIE1vbGVjdWxhciBTY2llbmNlIGFuZCBUZWNobm9sb2d5LCBBam91IFVuaXZlcnNp

dHksIFN1d29uIDQ0My03NDksIEtvcmVhLjwvYXV0aC1hZGRyZXNzPjx0aXRsZXM+PHRpdGxlPkdl

bmUgc2lsZW5jaW5nIGJ5IGNlbGwtcGVuZXRyYXRpbmcsIHNlcXVlbmNlLXNlbGVjdGl2ZSBhbmQg

bnVjbGVpYy1hY2lkIGh5ZHJvbHl6aW5nIGFudGlib2RpZXM8L3RpdGxlPjxzZWNvbmRhcnktdGl0

bGU+TnVjbGVpYyBBY2lkcyBSZXM8L3NlY29uZGFyeS10aXRsZT48YWx0LXRpdGxlPk51Y2xlaWMg

YWNpZHMgcmVzZWFyY2g8L2FsdC10aXRsZT48L3RpdGxlcz48cGVyaW9kaWNhbD48ZnVsbC10aXRs

ZT5OdWNsZWljIEFjaWRzIFJlczwvZnVsbC10aXRsZT48YWJici0xPk51Y2xlaWMgYWNpZHMgcmVz

ZWFyY2g8L2FiYnItMT48L3BlcmlvZGljYWw+PGFsdC1wZXJpb2RpY2FsPjxmdWxsLXRpdGxlPk51

Y2xlaWMgQWNpZHMgUmVzPC9mdWxsLXRpdGxlPjxhYmJyLTE+TnVjbGVpYyBhY2lkcyByZXNlYXJj

aDwvYWJici0xPjwvYWx0LXBlcmlvZGljYWw+PHBhZ2VzPjE1OTYtNjA5PC9wYWdlcz48dm9sdW1l

PjM4PC92b2x1bWU+PG51bWJlcj41PC9udW1iZXI+PGVkaXRpb24+MjAwOS8xMi8xNzwvZWRpdGlv

bj48a2V5d29yZHM+PGtleXdvcmQ+QW50aWJvZGllcywgQ2F0YWx5dGljL2NoZW1pc3RyeS8qbWV0

YWJvbGlzbTwva2V5d29yZD48a2V5d29yZD5BcG9wdG9zaXM8L2tleXdvcmQ+PGtleXdvcmQ+QmFz

ZSBTZXF1ZW5jZTwva2V5d29yZD48a2V5d29yZD5DZWxsIExpbmUsIFR1bW9yPC9rZXl3b3JkPjxr

ZXl3b3JkPkN5dG9zb2wvbWV0YWJvbGlzbTwva2V5d29yZD48a2V5d29yZD5ETkEsIFNpbmdsZS1T

dHJhbmRlZC9tZXRhYm9saXNtPC9rZXl3b3JkPjxrZXl3b3JkPkdlbmUgS25vY2tkb3duIFRlY2hu

aXF1ZXM8L2tleXdvcmQ+PGtleXdvcmQ+R2VuZSBMaWJyYXJ5PC9rZXl3b3JkPjxrZXl3b3JkPkh1

bWFuczwva2V5d29yZD48a2V5d29yZD5IeWRyb2x5c2lzPC9rZXl3b3JkPjxrZXl3b3JkPkltbXVu

b2dsb2J1bGluIExpZ2h0IENoYWlucy9jaGVtaXN0cnk8L2tleXdvcmQ+PGtleXdvcmQ+SW1tdW5v

Z2xvYnVsaW4gVmFyaWFibGUgUmVnaW9uL2NoZW1pc3RyeTwva2V5d29yZD48a2V5d29yZD4qUk5B

IEludGVyZmVyZW5jZTwva2V5d29yZD48a2V5d29yZD5STkEsIE1lc3Nlbmdlci8qbWV0YWJvbGlz

bTwva2V5d29yZD48a2V5d29yZD5SZWNlcHRvciwgZXJiQi0yL2FudGFnb25pc3RzICZhbXA7IGlu

aGliaXRvcnMvZ2VuZXRpY3MvbWV0YWJvbGlzbTwva2V5d29yZD48L2tleXdvcmRzPjxkYXRlcz48

eWVhcj4yMDEwPC95ZWFyPjxwdWItZGF0ZXM+PGRhdGU+TWFyPC9kYXRlPjwvcHViLWRhdGVzPjwv

ZGF0ZXM+PGlzYm4+MTM2Mi00OTYyIChFbGVjdHJvbmljKSYjeEQ7MDMwNS0xMDQ4IChMaW5raW5n

KTwvaXNibj48YWNjZXNzaW9uLW51bT4yMDAwNzYwMjwvYWNjZXNzaW9uLW51bT48d29yay10eXBl

PlJlc2VhcmNoIFN1cHBvcnQsIE5vbi1VLlMuIEdvdiZhcG9zO3Q8L3dvcmstdHlwZT48dXJscz48

cmVsYXRlZC11cmxzPjx1cmw+aHR0cDovL3d3dy5uY2JpLm5sbS5uaWguZ292L3B1Ym1lZC8yMDAw

NzYwMjwvdXJsPjwvcmVsYXRlZC11cmxzPjwvdXJscz48Y3VzdG9tMj4yODM2NTcyPC9jdXN0b20y

PjxlbGVjdHJvbmljLXJlc291cmNlLW51bT4xMC4xMDkzL25hci9na3AxMTQ1PC9lbGVjdHJvbmlj

LXJlc291cmNlLW51bT48bGFuZ3VhZ2U+ZW5nPC9sYW5ndWFnZT48L3JlY29yZD48L0NpdGU+PC9F

bmROb3RlPgB=

ADDIN EN.CITE PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5MZWU8L0F1dGhvcj48WWVhcj4yMDEwPC9ZZWFyPjxSZWNO

dW0+MjI8L1JlY051bT48RGlzcGxheVRleHQ+PHN0eWxlIGZhY2U9InN1cGVyc2NyaXB0Ij4zPC9z

dHlsZT48L0Rpc3BsYXlUZXh0PjxyZWNvcmQ+PHJlYy1udW1iZXI+MjI8L3JlYy1udW1iZXI+PGZv

cmVpZ24ta2V5cz48a2V5IGFwcD0iRU4iIGRiLWlkPSI1c3owZTUyenNzYXd4Y2VhMnhwcHc1NThm

cno1enpzMmQwMnYiPjIyPC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10eXBlIG5hbWU9IkpvdXJu

YWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1dGhvcnM+PGF1dGhvcj5M

ZWUsIFcuIFIuPC9hdXRob3I+PGF1dGhvcj5KYW5nLCBKLiBZLjwvYXV0aG9yPjxhdXRob3I+S2lt

LCBKLiBTLjwvYXV0aG9yPjxhdXRob3I+S3dvbiwgTS4gSC48L2F1dGhvcj48YXV0aG9yPktpbSwg

WS4gUy48L2F1dGhvcj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVzcz5EZXBh

cnRtZW50IG9mIE1vbGVjdWxhciBTY2llbmNlIGFuZCBUZWNobm9sb2d5LCBBam91IFVuaXZlcnNp

dHksIFN1d29uIDQ0My03NDksIEtvcmVhLjwvYXV0aC1hZGRyZXNzPjx0aXRsZXM+PHRpdGxlPkdl

bmUgc2lsZW5jaW5nIGJ5IGNlbGwtcGVuZXRyYXRpbmcsIHNlcXVlbmNlLXNlbGVjdGl2ZSBhbmQg

bnVjbGVpYy1hY2lkIGh5ZHJvbHl6aW5nIGFudGlib2RpZXM8L3RpdGxlPjxzZWNvbmRhcnktdGl0

bGU+TnVjbGVpYyBBY2lkcyBSZXM8L3NlY29uZGFyeS10aXRsZT48YWx0LXRpdGxlPk51Y2xlaWMg

YWNpZHMgcmVzZWFyY2g8L2FsdC10aXRsZT48L3RpdGxlcz48cGVyaW9kaWNhbD48ZnVsbC10aXRs

ZT5OdWNsZWljIEFjaWRzIFJlczwvZnVsbC10aXRsZT48YWJici0xPk51Y2xlaWMgYWNpZHMgcmVz

ZWFyY2g8L2FiYnItMT48L3BlcmlvZGljYWw+PGFsdC1wZXJpb2RpY2FsPjxmdWxsLXRpdGxlPk51

Y2xlaWMgQWNpZHMgUmVzPC9mdWxsLXRpdGxlPjxhYmJyLTE+TnVjbGVpYyBhY2lkcyByZXNlYXJj

aDwvYWJici0xPjwvYWx0LXBlcmlvZGljYWw+PHBhZ2VzPjE1OTYtNjA5PC9wYWdlcz48dm9sdW1l

PjM4PC92b2x1bWU+PG51bWJlcj41PC9udW1iZXI+PGVkaXRpb24+MjAwOS8xMi8xNzwvZWRpdGlv

bj48a2V5d29yZHM+PGtleXdvcmQ+QW50aWJvZGllcywgQ2F0YWx5dGljL2NoZW1pc3RyeS8qbWV0

YWJvbGlzbTwva2V5d29yZD48a2V5d29yZD5BcG9wdG9zaXM8L2tleXdvcmQ+PGtleXdvcmQ+QmFz

ZSBTZXF1ZW5jZTwva2V5d29yZD48a2V5d29yZD5DZWxsIExpbmUsIFR1bW9yPC9rZXl3b3JkPjxr

ZXl3b3JkPkN5dG9zb2wvbWV0YWJvbGlzbTwva2V5d29yZD48a2V5d29yZD5ETkEsIFNpbmdsZS1T

dHJhbmRlZC9tZXRhYm9saXNtPC9rZXl3b3JkPjxrZXl3b3JkPkdlbmUgS25vY2tkb3duIFRlY2hu

aXF1ZXM8L2tleXdvcmQ+PGtleXdvcmQ+R2VuZSBMaWJyYXJ5PC9rZXl3b3JkPjxrZXl3b3JkPkh1

bWFuczwva2V5d29yZD48a2V5d29yZD5IeWRyb2x5c2lzPC9rZXl3b3JkPjxrZXl3b3JkPkltbXVu

b2dsb2J1bGluIExpZ2h0IENoYWlucy9jaGVtaXN0cnk8L2tleXdvcmQ+PGtleXdvcmQ+SW1tdW5v

Z2xvYnVsaW4gVmFyaWFibGUgUmVnaW9uL2NoZW1pc3RyeTwva2V5d29yZD48a2V5d29yZD4qUk5B

IEludGVyZmVyZW5jZTwva2V5d29yZD48a2V5d29yZD5STkEsIE1lc3Nlbmdlci8qbWV0YWJvbGlz

bTwva2V5d29yZD48a2V5d29yZD5SZWNlcHRvciwgZXJiQi0yL2FudGFnb25pc3RzICZhbXA7IGlu

aGliaXRvcnMvZ2VuZXRpY3MvbWV0YWJvbGlzbTwva2V5d29yZD48L2tleXdvcmRzPjxkYXRlcz48

eWVhcj4yMDEwPC95ZWFyPjxwdWItZGF0ZXM+PGRhdGU+TWFyPC9kYXRlPjwvcHViLWRhdGVzPjwv

ZGF0ZXM+PGlzYm4+MTM2Mi00OTYyIChFbGVjdHJvbmljKSYjeEQ7MDMwNS0xMDQ4IChMaW5raW5n

KTwvaXNibj48YWNjZXNzaW9uLW51bT4yMDAwNzYwMjwvYWNjZXNzaW9uLW51bT48d29yay10eXBl

PlJlc2VhcmNoIFN1cHBvcnQsIE5vbi1VLlMuIEdvdiZhcG9zO3Q8L3dvcmstdHlwZT48dXJscz48

cmVsYXRlZC11cmxzPjx1cmw+aHR0cDovL3d3dy5uY2JpLm5sbS5uaWguZ292L3B1Ym1lZC8yMDAw

NzYwMjwvdXJsPjwvcmVsYXRlZC11cmxzPjwvdXJscz48Y3VzdG9tMj4yODM2NTcyPC9jdXN0b20y

PjxlbGVjdHJvbmljLXJlc291cmNlLW51bT4xMC4xMDkzL25hci9na3AxMTQ1PC9lbGVjdHJvbmlj

LXJlc291cmNlLW51bT48bGFuZ3VhZ2U+ZW5nPC9sYW5ndWFnZT48L3JlY29yZD48L0NpdGU+PC9F

bmROb3RlPgB=

ADDIN EN.CITE.DATA 3 Images show the merging of markers (green) and/or VLs (red) and Hochest33342-stained nuclei (blue) at the centered single confocal section. Image magnification, 630×; scale bar, 5 μm.Supplementary Figure S2. Generation of humanized hT4 VL with the highest frequent amino acids at the main VH-interacting positions on the consensus sequence of Vκ1 family germline segments. (A) Analysis of VH-VL interface of antibodies tested in our study. Left cartoon represents the VH-VL interface based on the three-dimensional structure of adalimumab (PDB 3wd5) PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5IdTwvQXV0aG9yPjxZZWFyPjIwMTM8L1llYXI+PFJlY051

bT4zODE8L1JlY051bT48RGlzcGxheVRleHQ+PHN0eWxlIGZhY2U9InN1cGVyc2NyaXB0Ij40PC9z

dHlsZT48L0Rpc3BsYXlUZXh0PjxyZWNvcmQ+PHJlYy1udW1iZXI+MzgxPC9yZWMtbnVtYmVyPjxm

b3JlaWduLWtleXM+PGtleSBhcHA9IkVOIiBkYi1pZD0iNXN6MGU1Mnpzc2F3eGNlYTJ4cHB3NTU4

ZnJ6NXp6czJkMDJ2Ij4zODE8L2tleT48L2ZvcmVpZ24ta2V5cz48cmVmLXR5cGUgbmFtZT0iSm91

cm5hbCBBcnRpY2xlIj4xNzwvcmVmLXR5cGU+PGNvbnRyaWJ1dG9ycz48YXV0aG9ycz48YXV0aG9y

Pkh1LCBTLjwvYXV0aG9yPjxhdXRob3I+TGlhbmcsIFMuPC9hdXRob3I+PGF1dGhvcj5HdW8sIEgu

PC9hdXRob3I+PGF1dGhvcj5aaGFuZywgRC48L2F1dGhvcj48YXV0aG9yPkxpLCBILjwvYXV0aG9y

PjxhdXRob3I+V2FuZywgWC48L2F1dGhvcj48YXV0aG9yPllhbmcsIFcuPC9hdXRob3I+PGF1dGhv

cj5RaWFuLCBXLjwvYXV0aG9yPjxhdXRob3I+SG91LCBTLjwvYXV0aG9yPjxhdXRob3I+V2FuZywg

SC48L2F1dGhvcj48YXV0aG9yPkd1bywgWS48L2F1dGhvcj48YXV0aG9yPkxvdSwgWi48L2F1dGhv

cj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVzcz5Gcm9tIHRoZSBMYWJvcmF0

b3J5IG9mIFN0cnVjdHVyYWwgQmlvbG9neSBhbmQgTWluaXN0cnkgb2YgRWR1Y2F0aW9uIExhYm9y

YXRvcnkgb2YgUHJvdGVpbiBTY2llbmNlLCBTY2hvb2wgb2YgTWVkaWNpbmUsIFRzaW5naHVhIFVu

aXZlcnNpdHksIEJlaWppbmcgMTAwMDg0LCBDaGluYS48L2F1dGgtYWRkcmVzcz48dGl0bGVzPjx0

aXRsZT5Db21wYXJpc29uIG9mIHRoZSBpbmhpYml0aW9uIG1lY2hhbmlzbXMgb2YgYWRhbGltdW1h

YiBhbmQgaW5mbGl4aW1hYiBpbiB0cmVhdGluZyB0dW1vciBuZWNyb3NpcyBmYWN0b3IgYWxwaGEt

YXNzb2NpYXRlZCBkaXNlYXNlcyBmcm9tIGEgbW9sZWN1bGFyIHZpZXc8L3RpdGxlPjxzZWNvbmRh

cnktdGl0bGU+SiBCaW9sIENoZW08L3NlY29uZGFyeS10aXRsZT48YWx0LXRpdGxlPlRoZSBKb3Vy

bmFsIG9mIGJpb2xvZ2ljYWwgY2hlbWlzdHJ5PC9hbHQtdGl0bGU+PC90aXRsZXM+PHBlcmlvZGlj

YWw+PGZ1bGwtdGl0bGU+SiBCaW9sIENoZW08L2Z1bGwtdGl0bGU+PGFiYnItMT5UaGUgSm91cm5h

bCBvZiBiaW9sb2dpY2FsIGNoZW1pc3RyeTwvYWJici0xPjwvcGVyaW9kaWNhbD48YWx0LXBlcmlv

ZGljYWw+PGZ1bGwtdGl0bGU+SiBCaW9sIENoZW08L2Z1bGwtdGl0bGU+PGFiYnItMT5UaGUgSm91

cm5hbCBvZiBiaW9sb2dpY2FsIGNoZW1pc3RyeTwvYWJici0xPjwvYWx0LXBlcmlvZGljYWw+PHBh

Z2VzPjI3MDU5LTY3PC9wYWdlcz48dm9sdW1lPjI4ODwvdm9sdW1lPjxudW1iZXI+Mzg8L251bWJl

cj48a2V5d29yZHM+PGtleXdvcmQ+QW50aS1JbmZsYW1tYXRvcnkgQWdlbnRzLCBOb24tU3Rlcm9p

ZGFsL2NoZW1pc3RyeS90aGVyYXBldXRpYyB1c2U8L2tleXdvcmQ+PGtleXdvcmQ+QW50aWJvZGll

cywgTW9ub2Nsb25hbC8qY2hlbWlzdHJ5L3RoZXJhcGV1dGljIHVzZTwva2V5d29yZD48a2V5d29y

ZD5BbnRpYm9kaWVzLCBNb25vY2xvbmFsLCBIdW1hbml6ZWQvKmNoZW1pc3RyeS90aGVyYXBldXRp

YyB1c2U8L2tleXdvcmQ+PGtleXdvcmQ+KkJpbmRpbmcgU2l0ZXMsIEFudGlib2R5PC9rZXl3b3Jk

PjxrZXl3b3JkPkNyeXN0YWxsb2dyYXBoeSwgWC1SYXk8L2tleXdvcmQ+PGtleXdvcmQ+SHVtYW5z

PC9rZXl3b3JkPjxrZXl3b3JkPipNb2RlbHMsIE1vbGVjdWxhcjwva2V5d29yZD48a2V5d29yZD5N

dXRhZ2VuZXNpczwva2V5d29yZD48a2V5d29yZD5Qcm90ZWluIFN0cnVjdHVyZSwgUXVhdGVybmFy

eTwva2V5d29yZD48a2V5d29yZD5Qcm90ZWluIFN0cnVjdHVyZSwgU2Vjb25kYXJ5PC9rZXl3b3Jk

PjxrZXl3b3JkPlR1bW9yIE5lY3Jvc2lzIEZhY3Rvci1hbHBoYS9hbnRhZ29uaXN0cyAmYW1wOyBp

bmhpYml0b3JzLypjaGVtaXN0cnkvZ2VuZXRpY3M8L2tleXdvcmQ+PC9rZXl3b3Jkcz48ZGF0ZXM+

PHllYXI+MjAxMzwveWVhcj48cHViLWRhdGVzPjxkYXRlPlNlcCAyMDwvZGF0ZT48L3B1Yi1kYXRl

cz48L2RhdGVzPjxpc2JuPjEwODMtMzUxWCAoRWxlY3Ryb25pYykmI3hEOzAwMjEtOTI1OCAoTGlu

a2luZyk8L2lzYm4+PGFjY2Vzc2lvbi1udW0+MjM5NDM2MTQ8L2FjY2Vzc2lvbi1udW0+PHVybHM+

PHJlbGF0ZWQtdXJscz48dXJsPmh0dHA6Ly93d3cubmNiaS5ubG0ubmloLmdvdi9wdWJtZWQvMjM5

NDM2MTQ8L3VybD48L3JlbGF0ZWQtdXJscz48L3VybHM+PGN1c3RvbTI+Mzc3OTcwNjwvY3VzdG9t

Mj48ZWxlY3Ryb25pYy1yZXNvdXJjZS1udW0+MTAuMTA3NC9qYmMuTTExMy40OTE1MzA8L2VsZWN0

cm9uaWMtcmVzb3VyY2UtbnVtPjwvcmVjb3JkPjwvQ2l0ZT48L0VuZE5vdGU+AG==

ADDIN EN.CITE PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5IdTwvQXV0aG9yPjxZZWFyPjIwMTM8L1llYXI+PFJlY051

bT4zODE8L1JlY051bT48RGlzcGxheVRleHQ+PHN0eWxlIGZhY2U9InN1cGVyc2NyaXB0Ij40PC9z

dHlsZT48L0Rpc3BsYXlUZXh0PjxyZWNvcmQ+PHJlYy1udW1iZXI+MzgxPC9yZWMtbnVtYmVyPjxm

b3JlaWduLWtleXM+PGtleSBhcHA9IkVOIiBkYi1pZD0iNXN6MGU1Mnpzc2F3eGNlYTJ4cHB3NTU4

ZnJ6NXp6czJkMDJ2Ij4zODE8L2tleT48L2ZvcmVpZ24ta2V5cz48cmVmLXR5cGUgbmFtZT0iSm91

cm5hbCBBcnRpY2xlIj4xNzwvcmVmLXR5cGU+PGNvbnRyaWJ1dG9ycz48YXV0aG9ycz48YXV0aG9y

Pkh1LCBTLjwvYXV0aG9yPjxhdXRob3I+TGlhbmcsIFMuPC9hdXRob3I+PGF1dGhvcj5HdW8sIEgu

PC9hdXRob3I+PGF1dGhvcj5aaGFuZywgRC48L2F1dGhvcj48YXV0aG9yPkxpLCBILjwvYXV0aG9y

PjxhdXRob3I+V2FuZywgWC48L2F1dGhvcj48YXV0aG9yPllhbmcsIFcuPC9hdXRob3I+PGF1dGhv

cj5RaWFuLCBXLjwvYXV0aG9yPjxhdXRob3I+SG91LCBTLjwvYXV0aG9yPjxhdXRob3I+V2FuZywg

SC48L2F1dGhvcj48YXV0aG9yPkd1bywgWS48L2F1dGhvcj48YXV0aG9yPkxvdSwgWi48L2F1dGhv

cj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVzcz5Gcm9tIHRoZSBMYWJvcmF0

b3J5IG9mIFN0cnVjdHVyYWwgQmlvbG9neSBhbmQgTWluaXN0cnkgb2YgRWR1Y2F0aW9uIExhYm9y

YXRvcnkgb2YgUHJvdGVpbiBTY2llbmNlLCBTY2hvb2wgb2YgTWVkaWNpbmUsIFRzaW5naHVhIFVu

aXZlcnNpdHksIEJlaWppbmcgMTAwMDg0LCBDaGluYS48L2F1dGgtYWRkcmVzcz48dGl0bGVzPjx0

aXRsZT5Db21wYXJpc29uIG9mIHRoZSBpbmhpYml0aW9uIG1lY2hhbmlzbXMgb2YgYWRhbGltdW1h

YiBhbmQgaW5mbGl4aW1hYiBpbiB0cmVhdGluZyB0dW1vciBuZWNyb3NpcyBmYWN0b3IgYWxwaGEt

YXNzb2NpYXRlZCBkaXNlYXNlcyBmcm9tIGEgbW9sZWN1bGFyIHZpZXc8L3RpdGxlPjxzZWNvbmRh

cnktdGl0bGU+SiBCaW9sIENoZW08L3NlY29uZGFyeS10aXRsZT48YWx0LXRpdGxlPlRoZSBKb3Vy

bmFsIG9mIGJpb2xvZ2ljYWwgY2hlbWlzdHJ5PC9hbHQtdGl0bGU+PC90aXRsZXM+PHBlcmlvZGlj

YWw+PGZ1bGwtdGl0bGU+SiBCaW9sIENoZW08L2Z1bGwtdGl0bGU+PGFiYnItMT5UaGUgSm91cm5h

bCBvZiBiaW9sb2dpY2FsIGNoZW1pc3RyeTwvYWJici0xPjwvcGVyaW9kaWNhbD48YWx0LXBlcmlv

ZGljYWw+PGZ1bGwtdGl0bGU+SiBCaW9sIENoZW08L2Z1bGwtdGl0bGU+PGFiYnItMT5UaGUgSm91

cm5hbCBvZiBiaW9sb2dpY2FsIGNoZW1pc3RyeTwvYWJici0xPjwvYWx0LXBlcmlvZGljYWw+PHBh

Z2VzPjI3MDU5LTY3PC9wYWdlcz48dm9sdW1lPjI4ODwvdm9sdW1lPjxudW1iZXI+Mzg8L251bWJl

cj48a2V5d29yZHM+PGtleXdvcmQ+QW50aS1JbmZsYW1tYXRvcnkgQWdlbnRzLCBOb24tU3Rlcm9p

ZGFsL2NoZW1pc3RyeS90aGVyYXBldXRpYyB1c2U8L2tleXdvcmQ+PGtleXdvcmQ+QW50aWJvZGll

cywgTW9ub2Nsb25hbC8qY2hlbWlzdHJ5L3RoZXJhcGV1dGljIHVzZTwva2V5d29yZD48a2V5d29y

ZD5BbnRpYm9kaWVzLCBNb25vY2xvbmFsLCBIdW1hbml6ZWQvKmNoZW1pc3RyeS90aGVyYXBldXRp

YyB1c2U8L2tleXdvcmQ+PGtleXdvcmQ+KkJpbmRpbmcgU2l0ZXMsIEFudGlib2R5PC9rZXl3b3Jk

PjxrZXl3b3JkPkNyeXN0YWxsb2dyYXBoeSwgWC1SYXk8L2tleXdvcmQ+PGtleXdvcmQ+SHVtYW5z

PC9rZXl3b3JkPjxrZXl3b3JkPipNb2RlbHMsIE1vbGVjdWxhcjwva2V5d29yZD48a2V5d29yZD5N

dXRhZ2VuZXNpczwva2V5d29yZD48a2V5d29yZD5Qcm90ZWluIFN0cnVjdHVyZSwgUXVhdGVybmFy

eTwva2V5d29yZD48a2V5d29yZD5Qcm90ZWluIFN0cnVjdHVyZSwgU2Vjb25kYXJ5PC9rZXl3b3Jk

PjxrZXl3b3JkPlR1bW9yIE5lY3Jvc2lzIEZhY3Rvci1hbHBoYS9hbnRhZ29uaXN0cyAmYW1wOyBp

bmhpYml0b3JzLypjaGVtaXN0cnkvZ2VuZXRpY3M8L2tleXdvcmQ+PC9rZXl3b3Jkcz48ZGF0ZXM+

PHllYXI+MjAxMzwveWVhcj48cHViLWRhdGVzPjxkYXRlPlNlcCAyMDwvZGF0ZT48L3B1Yi1kYXRl

cz48L2RhdGVzPjxpc2JuPjEwODMtMzUxWCAoRWxlY3Ryb25pYykmI3hEOzAwMjEtOTI1OCAoTGlu

a2luZyk8L2lzYm4+PGFjY2Vzc2lvbi1udW0+MjM5NDM2MTQ8L2FjY2Vzc2lvbi1udW0+PHVybHM+

PHJlbGF0ZWQtdXJscz48dXJsPmh0dHA6Ly93d3cubmNiaS5ubG0ubmloLmdvdi9wdWJtZWQvMjM5

NDM2MTQ8L3VybD48L3JlbGF0ZWQtdXJscz48L3VybHM+PGN1c3RvbTI+Mzc3OTcwNjwvY3VzdG9t

Mj48ZWxlY3Ryb25pYy1yZXNvdXJjZS1udW0+MTAuMTA3NC9qYmMuTTExMy40OTE1MzA8L2VsZWN0

cm9uaWMtcmVzb3VyY2UtbnVtPjwvcmVjb3JkPjwvQ2l0ZT48L0VuZE5vdGU+AG==

ADDIN EN.CITE.DATA 4, prepared by PyMol software. The right panel shows the 10 positions mostly involved in VH-VL interactions and the frequency of amino acids at each position in percentage (grey boxes), determined by structural analyses of 23 crystallized VH-VL interfaces. ADDIN EN.CITE <EndNote><Cite><Author>Vargas-Madrazo</Author><Year>2003</Year><RecNum>364</RecNum><DisplayText><style face="superscript">5</style></DisplayText><record><rec-number>364</rec-number><foreign-keys><key app="EN" db-id="5sz0e52zssawxcea2xppw558frz5zzs2d02v">364</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Vargas-Madrazo, E.</author><author>Paz-Garcia, E.</author></authors></contributors><auth-address>Instituto de Investigaciones Biologicas, Universidad Veracruzana, Xalapa, Veracruz, Mexico. evargas@uv.mx</auth-address><titles><title>An improved model of association for VH-VL immunoglobulin domains: asymmetries between VH and VL in the packing of some interface residues</title><secondary-title>J Mol Recognit</secondary-title></titles><periodical><full-title>J Mol Recognit</full-title></periodical><pages>113-20</pages><volume>16</volume><number>3</number><edition>2003/07/02</edition><keywords><keyword>Animals</keyword><keyword>Binding Sites, Antibody</keyword><keyword>Humans</keyword><keyword>Immunoglobulin Heavy Chains/*chemistry</keyword><keyword>Immunoglobulin Light Chains/*chemistry</keyword><keyword>Models, Molecular</keyword><keyword>Protein Conformation</keyword></keywords><dates><year>2003</year><pub-dates><date>May-Jun</date></pub-dates></dates><isbn>0952-3499 (Print)&#xD;0952-3499 (Linking)</isbn><accession-num>12833565</accession-num><urls><related-urls><url> Each position is color-coded based on the observed contact frequency from the analyses of 23 VH-VL interfaces. The interacting residues at the VH-VL interface with contact frequency above 50 % and between 20 % and 50 % are also indicated by red solid lines and black dotted lines, respectively. The amino acids at the 10 positions of VH and VL of antibodies (adalimumab, bevacizumab, TMab3 and TMab4) used in this study are also shown in single letter code. The hT3 VL conserved the highest frequent amino acids at all of the VH-interacting positions, except for two residues of K89 and S91 in VL-CDR3 that commonly interact with 103-3 and 95 residues in VH-CDR3, respectively. Thus we designed a hT3 variant, dubbed hT4, with two substitutions of K89Q and S91Y to have the most frequently used amino acids of human VLs at the two positions so that hT4 can be broadly associated with various human VH gene family. (B) Sequence alignment of VLs of hT3, hT4, adalimumab, and bevacizumab. The 10 positions of VL mostly involved in the interactions with VH are indicated by bold, blue color. CDRs indicated by brackets. Supplementary Figure S3. Biochemical characterizations of the purified parent mAbs and cytotransmabs. (A) Direct ELISA to determine the antigen binding activity of TMab4, HuT4, AvaT4, bevacizumab, and adalimumab antibodies (100 nM) to plate-coated antigen VEGF-A (0.05 ?g/ml) or TNFα (0.1 ?g/ml). (B) DNA hydrolyzing activity assay of the indicated antibodies by agarose gel electrophoresis. The supercoiled plasmid pUC19 (2.2 nM) was incubated with m3D8 scFv (100 nM, 500 nM) or IgG-format cytotransmabs (TMab4, HuT4, and AvaT4, 100 nM) for 1 h at 37 °C in TBS buffer (pH 7.4) containing 2 mM MgCl2 (indicated as ‘Mg’) or 50 mM EDTA (indicated as ‘E’). The reaction mixtures were analyzed by 0.7 % agarose gel electrophoresis, and then stained with ethidium bromide (EtBr). The plasmid incubated with the buffer alone and molecular mass markers were labeled as ‘Buffer’ and ‘M’, respectively. The m3D8 scFv was used as a positive control. We previously reported that m3D8 scFv hydrolyzes DNAs in the presence of Mg2+, but not EDTA.PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5KYW5nPC9BdXRob3I+PFllYXI+MjAwOTwvWWVhcj48UmVj

TnVtPjIwPC9SZWNOdW0+PERpc3BsYXlUZXh0PjxzdHlsZSBmYWNlPSJzdXBlcnNjcmlwdCI+Miwg

MywgNjwvc3R5bGU+PC9EaXNwbGF5VGV4dD48cmVjb3JkPjxyZWMtbnVtYmVyPjIwPC9yZWMtbnVt

YmVyPjxmb3JlaWduLWtleXM+PGtleSBhcHA9IkVOIiBkYi1pZD0icnpzZTB3c2Y5cnd4dzhldnph

bnBhdmY5YXp6OTJkd2RhdHRzIj4yMDwva2V5PjwvZm9yZWlnbi1rZXlzPjxyZWYtdHlwZSBuYW1l

PSJKb3VybmFsIEFydGljbGUiPjE3PC9yZWYtdHlwZT48Y29udHJpYnV0b3JzPjxhdXRob3JzPjxh

dXRob3I+SmFuZywgSi4gWS48L2F1dGhvcj48YXV0aG9yPkplb25nLCBKLiBHLjwvYXV0aG9yPjxh

dXRob3I+SnVuLCBILiBSLjwvYXV0aG9yPjxhdXRob3I+TGVlLCBTLiBDLjwvYXV0aG9yPjxhdXRo

b3I+S2ltLCBKLiBTLjwvYXV0aG9yPjxhdXRob3I+S2ltLCBZLiBTLjwvYXV0aG9yPjxhdXRob3I+

S3dvbiwgTS4gSC48L2F1dGhvcj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVz

cz5EZXBhcnRtZW50IG9mIE1pY3JvYmlvbG9neSwgQWpvdSBVbml2ZXJzaXR5IFNjaG9vbCBvZiBN

ZWRpY2luZSwgU3V3b24sIDQ0My03MjEsIEtvcmVhLjwvYXV0aC1hZGRyZXNzPjx0aXRsZXM+PHRp

dGxlPkEgbnVjbGVpYyBhY2lkLWh5ZHJvbHl6aW5nIGFudGlib2R5IHBlbmV0cmF0ZXMgaW50byBj

ZWxscyB2aWEgY2F2ZW9sYWUtbWVkaWF0ZWQgZW5kb2N5dG9zaXMsIGxvY2FsaXplcyBpbiB0aGUg

Y3l0b3NvbCBhbmQgZXhoaWJpdHMgY3l0b3RveGljaXR5PC90aXRsZT48c2Vjb25kYXJ5LXRpdGxl

PkNlbGwgTW9sIExpZmUgU2NpPC9zZWNvbmRhcnktdGl0bGU+PGFsdC10aXRsZT5DZWxsdWxhciBh

bmQgbW9sZWN1bGFyIGxpZmUgc2NpZW5jZXMgOiBDTUxTPC9hbHQtdGl0bGU+PC90aXRsZXM+PHBl

cmlvZGljYWw+PGZ1bGwtdGl0bGU+Q2VsbCBNb2wgTGlmZSBTY2k8L2Z1bGwtdGl0bGU+PGFiYnIt

MT5DZWxsdWxhciBhbmQgbW9sZWN1bGFyIGxpZmUgc2NpZW5jZXMgOiBDTUxTPC9hYmJyLTE+PC9w

ZXJpb2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVsbC10aXRsZT5DZWxsIE1vbCBMaWZlIFNjaTwv

ZnVsbC10aXRsZT48YWJici0xPkNlbGx1bGFyIGFuZCBtb2xlY3VsYXIgbGlmZSBzY2llbmNlcyA6

IENNTFM8L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4xOTg1LTk3PC9wYWdlcz48dm9s

dW1lPjY2PC92b2x1bWU+PG51bWJlcj4xMS0xMjwvbnVtYmVyPjxrZXl3b3Jkcz48a2V5d29yZD5B

bnRpYm9kaWVzLCBBbnRpbnVjbGVhci8qbWV0YWJvbGlzbS9waGFybWFjb2xvZ3k8L2tleXdvcmQ+

PGtleXdvcmQ+QXBvcHRvc2lzPC9rZXl3b3JkPjxrZXl3b3JkPkNhdmVvbGFlLypwaHlzaW9sb2d5

PC9rZXl3b3JkPjxrZXl3b3JkPkNlbGwgTWVtYnJhbmUgUGVybWVhYmlsaXR5PC9rZXl3b3JkPjxr

ZXl3b3JkPkNlbGwgU3Vydml2YWwvZHJ1ZyBlZmZlY3RzPC9rZXl3b3JkPjxrZXl3b3JkPkN5dG9z

b2wvKm1ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+RW5kb2N5dG9zaXM8L2tleXdvcmQ+PGtl

eXdvcmQ+SGVMYSBDZWxsczwva2V5d29yZD48a2V5d29yZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdv

cmQ+SHlkcm9seXNpczwva2V5d29yZD48a2V5d29yZD5JbW11bm9nbG9idWxpbiBWYXJpYWJsZSBS

ZWdpb24vKm1ldGFib2xpc20vcGhhcm1hY29sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPk1lbWJyYW5l

IE1pY3JvZG9tYWluczwva2V5d29yZD48a2V5d29yZD5Qcm90ZW9nbHljYW5zL21ldGFib2xpc208

L2tleXdvcmQ+PGtleXdvcmQ+Uk5BL21ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+Umlib251

Y2xlYXNlcy9tZXRhYm9saXNtPC9rZXl3b3JkPjwva2V5d29yZHM+PGRhdGVzPjx5ZWFyPjIwMDk8

L3llYXI+PHB1Yi1kYXRlcz48ZGF0ZT5KdW48L2RhdGU+PC9wdWItZGF0ZXM+PC9kYXRlcz48aXNi

bj4xNDIwLTkwNzEgKEVsZWN0cm9uaWMpJiN4RDsxNDIwLTY4MlggKExpbmtpbmcpPC9pc2JuPjxh

Y2Nlc3Npb24tbnVtPjE5MzczNDM0PC9hY2Nlc3Npb24tbnVtPjx1cmxzPjxyZWxhdGVkLXVybHM+

PHVybD5odHRwOi8vd3d3Lm5jYmkubmxtLm5paC5nb3YvcHVibWVkLzE5MzczNDM0PC91cmw+PC9y

ZWxhdGVkLXVybHM+PC91cmxzPjxlbGVjdHJvbmljLXJlc291cmNlLW51bT4xMC4xMDA3L3MwMDAx

OC0wMDktOTE3OS0yPC9lbGVjdHJvbmljLXJlc291cmNlLW51bT48L3JlY29yZD48L0NpdGU+PENp

dGU+PEF1dGhvcj5LaW08L0F1dGhvcj48WWVhcj4yMDA5PC9ZZWFyPjxSZWNOdW0+NDU8L1JlY051

bT48cmVjb3JkPjxyZWMtbnVtYmVyPjQ1PC9yZWMtbnVtYmVyPjxmb3JlaWduLWtleXM+PGtleSBh

cHA9IkVOIiBkYi1pZD0iNXN6MGU1Mnpzc2F3eGNlYTJ4cHB3NTU4ZnJ6NXp6czJkMDJ2Ij40NTwv

a2V5PjwvZm9yZWlnbi1rZXlzPjxyZWYtdHlwZSBuYW1lPSJKb3VybmFsIEFydGljbGUiPjE3PC9y

ZWYtdHlwZT48Y29udHJpYnV0b3JzPjxhdXRob3JzPjxhdXRob3I+S2ltLCBELiBTLjwvYXV0aG9y

PjxhdXRob3I+TGVlLCBTLiBILjwvYXV0aG9yPjxhdXRob3I+S2ltLCBKLiBTLjwvYXV0aG9yPjxh

dXRob3I+TGVlLCBTLiBDLjwvYXV0aG9yPjxhdXRob3I+S3dvbiwgTS4gSC48L2F1dGhvcj48YXV0

aG9yPktpbSwgWS4gUy48L2F1dGhvcj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRk

cmVzcz5EZXBhcnRtZW50IG9mIE1vbGVjdWxhciBTY2llbmNlIGFuZCBUZWNobm9sb2d5LCBBam91

IFVuaXZlcnNpdHksIFNhbiA1LCBXb25jaGVvbi1kb25nLCBZZW9uZ3RvbmctZ3UsIFN1d29uIDQ0

My03NDksIFJlcHVibGljIG9mIEtvcmVhLjwvYXV0aC1hZGRyZXNzPjx0aXRsZXM+PHRpdGxlPkdl

bmVyYXRpb24gb2YgaHVtYW5pemVkIGFudGktRE5BIGh5ZHJvbHl6aW5nIGNhdGFseXRpYyBhbnRp

Ym9kaWVzIGJ5IGNvbXBsZW1lbnRhcml0eSBkZXRlcm1pbmluZyByZWdpb24gZ3JhZnRpbmc8L3Rp

dGxlPjxzZWNvbmRhcnktdGl0bGU+QmlvY2hlbSBCaW9waHlzIFJlcyBDb21tdW48L3NlY29uZGFy

eS10aXRsZT48L3RpdGxlcz48cGVyaW9kaWNhbD48ZnVsbC10aXRsZT5CaW9jaGVtIEJpb3BoeXMg

UmVzIENvbW11bjwvZnVsbC10aXRsZT48YWJici0xPkJpb2NoZW1pY2FsIGFuZCBiaW9waHlzaWNh

bCByZXNlYXJjaCBjb21tdW5pY2F0aW9uczwvYWJici0xPjwvcGVyaW9kaWNhbD48cGFnZXM+MzE0

LTg8L3BhZ2VzPjx2b2x1bWU+Mzc5PC92b2x1bWU+PG51bWJlcj4yPC9udW1iZXI+PGRhdGVzPjx5

ZWFyPjIwMDk8L3llYXI+PHB1Yi1kYXRlcz48ZGF0ZT5GZWIgNjwvZGF0ZT48L3B1Yi1kYXRlcz48

L2RhdGVzPjxhY2Nlc3Npb24tbnVtPjE5MTAzMTcxPC9hY2Nlc3Npb24tbnVtPjx1cmxzPjxyZWxh

dGVkLXVybHM+PHVybD5odHRwOi8vd3d3Lm5jYmkubmxtLm5paC5nb3YvZW50cmV6L3F1ZXJ5LmZj

Z2k/Y21kPVJldHJpZXZlJmFtcDtkYj1QdWJNZWQmYW1wO2RvcHQ9Q2l0YXRpb24mYW1wO2xpc3Rf

dWlkcz0xOTEwMzE3MSA8L3VybD48L3JlbGF0ZWQtdXJscz48L3VybHM+PC9yZWNvcmQ+PC9DaXRl

PjxDaXRlPjxBdXRob3I+TGVlPC9BdXRob3I+PFllYXI+MjAxMDwvWWVhcj48UmVjTnVtPjIyPC9S

ZWNOdW0+PHJlY29yZD48cmVjLW51bWJlcj4yMjwvcmVjLW51bWJlcj48Zm9yZWlnbi1rZXlzPjxr

ZXkgYXBwPSJFTiIgZGItaWQ9IjVzejBlNTJ6c3Nhd3hjZWEyeHBwdzU1OGZyejV6enMyZDAydiI+

MjI8L2tleT48L2ZvcmVpZ24ta2V5cz48cmVmLXR5cGUgbmFtZT0iSm91cm5hbCBBcnRpY2xlIj4x

NzwvcmVmLXR5cGU+PGNvbnRyaWJ1dG9ycz48YXV0aG9ycz48YXV0aG9yPkxlZSwgVy4gUi48L2F1

dGhvcj48YXV0aG9yPkphbmcsIEouIFkuPC9hdXRob3I+PGF1dGhvcj5LaW0sIEouIFMuPC9hdXRo

b3I+PGF1dGhvcj5Ld29uLCBNLiBILjwvYXV0aG9yPjxhdXRob3I+S2ltLCBZLiBTLjwvYXV0aG9y

PjwvYXV0aG9ycz48L2NvbnRyaWJ1dG9ycz48YXV0aC1hZGRyZXNzPkRlcGFydG1lbnQgb2YgTW9s

ZWN1bGFyIFNjaWVuY2UgYW5kIFRlY2hub2xvZ3ksIEFqb3UgVW5pdmVyc2l0eSwgU3V3b24gNDQz

LTc0OSwgS29yZWEuPC9hdXRoLWFkZHJlc3M+PHRpdGxlcz48dGl0bGU+R2VuZSBzaWxlbmNpbmcg

YnkgY2VsbC1wZW5ldHJhdGluZywgc2VxdWVuY2Utc2VsZWN0aXZlIGFuZCBudWNsZWljLWFjaWQg

aHlkcm9seXppbmcgYW50aWJvZGllczwvdGl0bGU+PHNlY29uZGFyeS10aXRsZT5OdWNsZWljIEFj

aWRzIFJlczwvc2Vjb25kYXJ5LXRpdGxlPjxhbHQtdGl0bGU+TnVjbGVpYyBhY2lkcyByZXNlYXJj

aDwvYWx0LXRpdGxlPjwvdGl0bGVzPjxwZXJpb2RpY2FsPjxmdWxsLXRpdGxlPk51Y2xlaWMgQWNp

ZHMgUmVzPC9mdWxsLXRpdGxlPjxhYmJyLTE+TnVjbGVpYyBhY2lkcyByZXNlYXJjaDwvYWJici0x

PjwvcGVyaW9kaWNhbD48YWx0LXBlcmlvZGljYWw+PGZ1bGwtdGl0bGU+TnVjbGVpYyBBY2lkcyBS

ZXM8L2Z1bGwtdGl0bGU+PGFiYnItMT5OdWNsZWljIGFjaWRzIHJlc2VhcmNoPC9hYmJyLTE+PC9h

bHQtcGVyaW9kaWNhbD48cGFnZXM+MTU5Ni02MDk8L3BhZ2VzPjx2b2x1bWU+Mzg8L3ZvbHVtZT48

bnVtYmVyPjU8L251bWJlcj48ZWRpdGlvbj4yMDA5LzEyLzE3PC9lZGl0aW9uPjxrZXl3b3Jkcz48

a2V5d29yZD5BbnRpYm9kaWVzLCBDYXRhbHl0aWMvY2hlbWlzdHJ5LyptZXRhYm9saXNtPC9rZXl3

b3JkPjxrZXl3b3JkPkFwb3B0b3Npczwva2V5d29yZD48a2V5d29yZD5CYXNlIFNlcXVlbmNlPC9r

ZXl3b3JkPjxrZXl3b3JkPkNlbGwgTGluZSwgVHVtb3I8L2tleXdvcmQ+PGtleXdvcmQ+Q3l0b3Nv

bC9tZXRhYm9saXNtPC9rZXl3b3JkPjxrZXl3b3JkPkROQSwgU2luZ2xlLVN0cmFuZGVkL21ldGFi

b2xpc208L2tleXdvcmQ+PGtleXdvcmQ+R2VuZSBLbm9ja2Rvd24gVGVjaG5pcXVlczwva2V5d29y

ZD48a2V5d29yZD5HZW5lIExpYnJhcnk8L2tleXdvcmQ+PGtleXdvcmQ+SHVtYW5zPC9rZXl3b3Jk

PjxrZXl3b3JkPkh5ZHJvbHlzaXM8L2tleXdvcmQ+PGtleXdvcmQ+SW1tdW5vZ2xvYnVsaW4gTGln

aHQgQ2hhaW5zL2NoZW1pc3RyeTwva2V5d29yZD48a2V5d29yZD5JbW11bm9nbG9idWxpbiBWYXJp

YWJsZSBSZWdpb24vY2hlbWlzdHJ5PC9rZXl3b3JkPjxrZXl3b3JkPipSTkEgSW50ZXJmZXJlbmNl

PC9rZXl3b3JkPjxrZXl3b3JkPlJOQSwgTWVzc2VuZ2VyLyptZXRhYm9saXNtPC9rZXl3b3JkPjxr

ZXl3b3JkPlJlY2VwdG9yLCBlcmJCLTIvYW50YWdvbmlzdHMgJmFtcDsgaW5oaWJpdG9ycy9nZW5l

dGljcy9tZXRhYm9saXNtPC9rZXl3b3JkPjwva2V5d29yZHM+PGRhdGVzPjx5ZWFyPjIwMTA8L3ll

YXI+PHB1Yi1kYXRlcz48ZGF0ZT5NYXI8L2RhdGU+PC9wdWItZGF0ZXM+PC9kYXRlcz48aXNibj4x

MzYyLTQ5NjIgKEVsZWN0cm9uaWMpJiN4RDswMzA1LTEwNDggKExpbmtpbmcpPC9pc2JuPjxhY2Nl

c3Npb24tbnVtPjIwMDA3NjAyPC9hY2Nlc3Npb24tbnVtPjx3b3JrLXR5cGU+UmVzZWFyY2ggU3Vw

cG9ydCwgTm9uLVUuUy4gR292JmFwb3M7dDwvd29yay10eXBlPjx1cmxzPjxyZWxhdGVkLXVybHM+

PHVybD5odHRwOi8vd3d3Lm5jYmkubmxtLm5paC5nb3YvcHVibWVkLzIwMDA3NjAyPC91cmw+PC9y

ZWxhdGVkLXVybHM+PC91cmxzPjxjdXN0b20yPjI4MzY1NzI8L2N1c3RvbTI+PGVsZWN0cm9uaWMt

cmVzb3VyY2UtbnVtPjEwLjEwOTMvbmFyL2drcDExNDU8L2VsZWN0cm9uaWMtcmVzb3VyY2UtbnVt

PjxsYW5ndWFnZT5lbmc8L2xhbmd1YWdlPjwvcmVjb3JkPjwvQ2l0ZT48L0VuZE5vdGU+

ADDIN EN.CITE PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5KYW5nPC9BdXRob3I+PFllYXI+MjAwOTwvWWVhcj48UmVj

TnVtPjIwPC9SZWNOdW0+PERpc3BsYXlUZXh0PjxzdHlsZSBmYWNlPSJzdXBlcnNjcmlwdCI+Miwg

MywgNjwvc3R5bGU+PC9EaXNwbGF5VGV4dD48cmVjb3JkPjxyZWMtbnVtYmVyPjIwPC9yZWMtbnVt

YmVyPjxmb3JlaWduLWtleXM+PGtleSBhcHA9IkVOIiBkYi1pZD0icnpzZTB3c2Y5cnd4dzhldnph

bnBhdmY5YXp6OTJkd2RhdHRzIj4yMDwva2V5PjwvZm9yZWlnbi1rZXlzPjxyZWYtdHlwZSBuYW1l

PSJKb3VybmFsIEFydGljbGUiPjE3PC9yZWYtdHlwZT48Y29udHJpYnV0b3JzPjxhdXRob3JzPjxh

dXRob3I+SmFuZywgSi4gWS48L2F1dGhvcj48YXV0aG9yPkplb25nLCBKLiBHLjwvYXV0aG9yPjxh

dXRob3I+SnVuLCBILiBSLjwvYXV0aG9yPjxhdXRob3I+TGVlLCBTLiBDLjwvYXV0aG9yPjxhdXRo

b3I+S2ltLCBKLiBTLjwvYXV0aG9yPjxhdXRob3I+S2ltLCBZLiBTLjwvYXV0aG9yPjxhdXRob3I+

S3dvbiwgTS4gSC48L2F1dGhvcj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVz

cz5EZXBhcnRtZW50IG9mIE1pY3JvYmlvbG9neSwgQWpvdSBVbml2ZXJzaXR5IFNjaG9vbCBvZiBN

ZWRpY2luZSwgU3V3b24sIDQ0My03MjEsIEtvcmVhLjwvYXV0aC1hZGRyZXNzPjx0aXRsZXM+PHRp

dGxlPkEgbnVjbGVpYyBhY2lkLWh5ZHJvbHl6aW5nIGFudGlib2R5IHBlbmV0cmF0ZXMgaW50byBj

ZWxscyB2aWEgY2F2ZW9sYWUtbWVkaWF0ZWQgZW5kb2N5dG9zaXMsIGxvY2FsaXplcyBpbiB0aGUg

Y3l0b3NvbCBhbmQgZXhoaWJpdHMgY3l0b3RveGljaXR5PC90aXRsZT48c2Vjb25kYXJ5LXRpdGxl

PkNlbGwgTW9sIExpZmUgU2NpPC9zZWNvbmRhcnktdGl0bGU+PGFsdC10aXRsZT5DZWxsdWxhciBh

bmQgbW9sZWN1bGFyIGxpZmUgc2NpZW5jZXMgOiBDTUxTPC9hbHQtdGl0bGU+PC90aXRsZXM+PHBl

cmlvZGljYWw+PGZ1bGwtdGl0bGU+Q2VsbCBNb2wgTGlmZSBTY2k8L2Z1bGwtdGl0bGU+PGFiYnIt

MT5DZWxsdWxhciBhbmQgbW9sZWN1bGFyIGxpZmUgc2NpZW5jZXMgOiBDTUxTPC9hYmJyLTE+PC9w

ZXJpb2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVsbC10aXRsZT5DZWxsIE1vbCBMaWZlIFNjaTwv

ZnVsbC10aXRsZT48YWJici0xPkNlbGx1bGFyIGFuZCBtb2xlY3VsYXIgbGlmZSBzY2llbmNlcyA6

IENNTFM8L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4xOTg1LTk3PC9wYWdlcz48dm9s

dW1lPjY2PC92b2x1bWU+PG51bWJlcj4xMS0xMjwvbnVtYmVyPjxrZXl3b3Jkcz48a2V5d29yZD5B

bnRpYm9kaWVzLCBBbnRpbnVjbGVhci8qbWV0YWJvbGlzbS9waGFybWFjb2xvZ3k8L2tleXdvcmQ+

PGtleXdvcmQ+QXBvcHRvc2lzPC9rZXl3b3JkPjxrZXl3b3JkPkNhdmVvbGFlLypwaHlzaW9sb2d5

PC9rZXl3b3JkPjxrZXl3b3JkPkNlbGwgTWVtYnJhbmUgUGVybWVhYmlsaXR5PC9rZXl3b3JkPjxr

ZXl3b3JkPkNlbGwgU3Vydml2YWwvZHJ1ZyBlZmZlY3RzPC9rZXl3b3JkPjxrZXl3b3JkPkN5dG9z

b2wvKm1ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+RW5kb2N5dG9zaXM8L2tleXdvcmQ+PGtl

eXdvcmQ+SGVMYSBDZWxsczwva2V5d29yZD48a2V5d29yZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdv

cmQ+SHlkcm9seXNpczwva2V5d29yZD48a2V5d29yZD5JbW11bm9nbG9idWxpbiBWYXJpYWJsZSBS

ZWdpb24vKm1ldGFib2xpc20vcGhhcm1hY29sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPk1lbWJyYW5l

IE1pY3JvZG9tYWluczwva2V5d29yZD48a2V5d29yZD5Qcm90ZW9nbHljYW5zL21ldGFib2xpc208

L2tleXdvcmQ+PGtleXdvcmQ+Uk5BL21ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+Umlib251

Y2xlYXNlcy9tZXRhYm9saXNtPC9rZXl3b3JkPjwva2V5d29yZHM+PGRhdGVzPjx5ZWFyPjIwMDk8

L3llYXI+PHB1Yi1kYXRlcz48ZGF0ZT5KdW48L2RhdGU+PC9wdWItZGF0ZXM+PC9kYXRlcz48aXNi

bj4xNDIwLTkwNzEgKEVsZWN0cm9uaWMpJiN4RDsxNDIwLTY4MlggKExpbmtpbmcpPC9pc2JuPjxh

Y2Nlc3Npb24tbnVtPjE5MzczNDM0PC9hY2Nlc3Npb24tbnVtPjx1cmxzPjxyZWxhdGVkLXVybHM+

PHVybD5odHRwOi8vd3d3Lm5jYmkubmxtLm5paC5nb3YvcHVibWVkLzE5MzczNDM0PC91cmw+PC9y

ZWxhdGVkLXVybHM+PC91cmxzPjxlbGVjdHJvbmljLXJlc291cmNlLW51bT4xMC4xMDA3L3MwMDAx

OC0wMDktOTE3OS0yPC9lbGVjdHJvbmljLXJlc291cmNlLW51bT48L3JlY29yZD48L0NpdGU+PENp

dGU+PEF1dGhvcj5LaW08L0F1dGhvcj48WWVhcj4yMDA5PC9ZZWFyPjxSZWNOdW0+NDU8L1JlY051

bT48cmVjb3JkPjxyZWMtbnVtYmVyPjQ1PC9yZWMtbnVtYmVyPjxmb3JlaWduLWtleXM+PGtleSBh

cHA9IkVOIiBkYi1pZD0iNXN6MGU1Mnpzc2F3eGNlYTJ4cHB3NTU4ZnJ6NXp6czJkMDJ2Ij40NTwv

a2V5PjwvZm9yZWlnbi1rZXlzPjxyZWYtdHlwZSBuYW1lPSJKb3VybmFsIEFydGljbGUiPjE3PC9y

ZWYtdHlwZT48Y29udHJpYnV0b3JzPjxhdXRob3JzPjxhdXRob3I+S2ltLCBELiBTLjwvYXV0aG9y

PjxhdXRob3I+TGVlLCBTLiBILjwvYXV0aG9yPjxhdXRob3I+S2ltLCBKLiBTLjwvYXV0aG9yPjxh

dXRob3I+TGVlLCBTLiBDLjwvYXV0aG9yPjxhdXRob3I+S3dvbiwgTS4gSC48L2F1dGhvcj48YXV0

aG9yPktpbSwgWS4gUy48L2F1dGhvcj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRk

cmVzcz5EZXBhcnRtZW50IG9mIE1vbGVjdWxhciBTY2llbmNlIGFuZCBUZWNobm9sb2d5LCBBam91

IFVuaXZlcnNpdHksIFNhbiA1LCBXb25jaGVvbi1kb25nLCBZZW9uZ3RvbmctZ3UsIFN1d29uIDQ0

My03NDksIFJlcHVibGljIG9mIEtvcmVhLjwvYXV0aC1hZGRyZXNzPjx0aXRsZXM+PHRpdGxlPkdl

bmVyYXRpb24gb2YgaHVtYW5pemVkIGFudGktRE5BIGh5ZHJvbHl6aW5nIGNhdGFseXRpYyBhbnRp

Ym9kaWVzIGJ5IGNvbXBsZW1lbnRhcml0eSBkZXRlcm1pbmluZyByZWdpb24gZ3JhZnRpbmc8L3Rp

dGxlPjxzZWNvbmRhcnktdGl0bGU+QmlvY2hlbSBCaW9waHlzIFJlcyBDb21tdW48L3NlY29uZGFy

eS10aXRsZT48L3RpdGxlcz48cGVyaW9kaWNhbD48ZnVsbC10aXRsZT5CaW9jaGVtIEJpb3BoeXMg

UmVzIENvbW11bjwvZnVsbC10aXRsZT48YWJici0xPkJpb2NoZW1pY2FsIGFuZCBiaW9waHlzaWNh

bCByZXNlYXJjaCBjb21tdW5pY2F0aW9uczwvYWJici0xPjwvcGVyaW9kaWNhbD48cGFnZXM+MzE0

LTg8L3BhZ2VzPjx2b2x1bWU+Mzc5PC92b2x1bWU+PG51bWJlcj4yPC9udW1iZXI+PGRhdGVzPjx5

ZWFyPjIwMDk8L3llYXI+PHB1Yi1kYXRlcz48ZGF0ZT5GZWIgNjwvZGF0ZT48L3B1Yi1kYXRlcz48

L2RhdGVzPjxhY2Nlc3Npb24tbnVtPjE5MTAzMTcxPC9hY2Nlc3Npb24tbnVtPjx1cmxzPjxyZWxh

dGVkLXVybHM+PHVybD5odHRwOi8vd3d3Lm5jYmkubmxtLm5paC5nb3YvZW50cmV6L3F1ZXJ5LmZj

Z2k/Y21kPVJldHJpZXZlJmFtcDtkYj1QdWJNZWQmYW1wO2RvcHQ9Q2l0YXRpb24mYW1wO2xpc3Rf

dWlkcz0xOTEwMzE3MSA8L3VybD48L3JlbGF0ZWQtdXJscz48L3VybHM+PC9yZWNvcmQ+PC9DaXRl

PjxDaXRlPjxBdXRob3I+TGVlPC9BdXRob3I+PFllYXI+MjAxMDwvWWVhcj48UmVjTnVtPjIyPC9S

ZWNOdW0+PHJlY29yZD48cmVjLW51bWJlcj4yMjwvcmVjLW51bWJlcj48Zm9yZWlnbi1rZXlzPjxr

ZXkgYXBwPSJFTiIgZGItaWQ9IjVzejBlNTJ6c3Nhd3hjZWEyeHBwdzU1OGZyejV6enMyZDAydiI+

MjI8L2tleT48L2ZvcmVpZ24ta2V5cz48cmVmLXR5cGUgbmFtZT0iSm91cm5hbCBBcnRpY2xlIj4x

NzwvcmVmLXR5cGU+PGNvbnRyaWJ1dG9ycz48YXV0aG9ycz48YXV0aG9yPkxlZSwgVy4gUi48L2F1

dGhvcj48YXV0aG9yPkphbmcsIEouIFkuPC9hdXRob3I+PGF1dGhvcj5LaW0sIEouIFMuPC9hdXRo

b3I+PGF1dGhvcj5Ld29uLCBNLiBILjwvYXV0aG9yPjxhdXRob3I+S2ltLCBZLiBTLjwvYXV0aG9y

PjwvYXV0aG9ycz48L2NvbnRyaWJ1dG9ycz48YXV0aC1hZGRyZXNzPkRlcGFydG1lbnQgb2YgTW9s

ZWN1bGFyIFNjaWVuY2UgYW5kIFRlY2hub2xvZ3ksIEFqb3UgVW5pdmVyc2l0eSwgU3V3b24gNDQz

LTc0OSwgS29yZWEuPC9hdXRoLWFkZHJlc3M+PHRpdGxlcz48dGl0bGU+R2VuZSBzaWxlbmNpbmcg

YnkgY2VsbC1wZW5ldHJhdGluZywgc2VxdWVuY2Utc2VsZWN0aXZlIGFuZCBudWNsZWljLWFjaWQg

aHlkcm9seXppbmcgYW50aWJvZGllczwvdGl0bGU+PHNlY29uZGFyeS10aXRsZT5OdWNsZWljIEFj

aWRzIFJlczwvc2Vjb25kYXJ5LXRpdGxlPjxhbHQtdGl0bGU+TnVjbGVpYyBhY2lkcyByZXNlYXJj

aDwvYWx0LXRpdGxlPjwvdGl0bGVzPjxwZXJpb2RpY2FsPjxmdWxsLXRpdGxlPk51Y2xlaWMgQWNp

ZHMgUmVzPC9mdWxsLXRpdGxlPjxhYmJyLTE+TnVjbGVpYyBhY2lkcyByZXNlYXJjaDwvYWJici0x

PjwvcGVyaW9kaWNhbD48YWx0LXBlcmlvZGljYWw+PGZ1bGwtdGl0bGU+TnVjbGVpYyBBY2lkcyBS

ZXM8L2Z1bGwtdGl0bGU+PGFiYnItMT5OdWNsZWljIGFjaWRzIHJlc2VhcmNoPC9hYmJyLTE+PC9h

bHQtcGVyaW9kaWNhbD48cGFnZXM+MTU5Ni02MDk8L3BhZ2VzPjx2b2x1bWU+Mzg8L3ZvbHVtZT48

bnVtYmVyPjU8L251bWJlcj48ZWRpdGlvbj4yMDA5LzEyLzE3PC9lZGl0aW9uPjxrZXl3b3Jkcz48

a2V5d29yZD5BbnRpYm9kaWVzLCBDYXRhbHl0aWMvY2hlbWlzdHJ5LyptZXRhYm9saXNtPC9rZXl3

b3JkPjxrZXl3b3JkPkFwb3B0b3Npczwva2V5d29yZD48a2V5d29yZD5CYXNlIFNlcXVlbmNlPC9r

ZXl3b3JkPjxrZXl3b3JkPkNlbGwgTGluZSwgVHVtb3I8L2tleXdvcmQ+PGtleXdvcmQ+Q3l0b3Nv

bC9tZXRhYm9saXNtPC9rZXl3b3JkPjxrZXl3b3JkPkROQSwgU2luZ2xlLVN0cmFuZGVkL21ldGFi

b2xpc208L2tleXdvcmQ+PGtleXdvcmQ+R2VuZSBLbm9ja2Rvd24gVGVjaG5pcXVlczwva2V5d29y

ZD48a2V5d29yZD5HZW5lIExpYnJhcnk8L2tleXdvcmQ+PGtleXdvcmQ+SHVtYW5zPC9rZXl3b3Jk

PjxrZXl3b3JkPkh5ZHJvbHlzaXM8L2tleXdvcmQ+PGtleXdvcmQ+SW1tdW5vZ2xvYnVsaW4gTGln

aHQgQ2hhaW5zL2NoZW1pc3RyeTwva2V5d29yZD48a2V5d29yZD5JbW11bm9nbG9idWxpbiBWYXJp

YWJsZSBSZWdpb24vY2hlbWlzdHJ5PC9rZXl3b3JkPjxrZXl3b3JkPipSTkEgSW50ZXJmZXJlbmNl

PC9rZXl3b3JkPjxrZXl3b3JkPlJOQSwgTWVzc2VuZ2VyLyptZXRhYm9saXNtPC9rZXl3b3JkPjxr

ZXl3b3JkPlJlY2VwdG9yLCBlcmJCLTIvYW50YWdvbmlzdHMgJmFtcDsgaW5oaWJpdG9ycy9nZW5l

dGljcy9tZXRhYm9saXNtPC9rZXl3b3JkPjwva2V5d29yZHM+PGRhdGVzPjx5ZWFyPjIwMTA8L3ll

YXI+PHB1Yi1kYXRlcz48ZGF0ZT5NYXI8L2RhdGU+PC9wdWItZGF0ZXM+PC9kYXRlcz48aXNibj4x

MzYyLTQ5NjIgKEVsZWN0cm9uaWMpJiN4RDswMzA1LTEwNDggKExpbmtpbmcpPC9pc2JuPjxhY2Nl

c3Npb24tbnVtPjIwMDA3NjAyPC9hY2Nlc3Npb24tbnVtPjx3b3JrLXR5cGU+UmVzZWFyY2ggU3Vw

cG9ydCwgTm9uLVUuUy4gR292JmFwb3M7dDwvd29yay10eXBlPjx1cmxzPjxyZWxhdGVkLXVybHM+

PHVybD5odHRwOi8vd3d3Lm5jYmkubmxtLm5paC5nb3YvcHVibWVkLzIwMDA3NjAyPC91cmw+PC9y

ZWxhdGVkLXVybHM+PC91cmxzPjxjdXN0b20yPjI4MzY1NzI8L2N1c3RvbTI+PGVsZWN0cm9uaWMt

cmVzb3VyY2UtbnVtPjEwLjEwOTMvbmFyL2drcDExNDU8L2VsZWN0cm9uaWMtcmVzb3VyY2UtbnVt

PjxsYW5ndWFnZT5lbmc8L2xhbmd1YWdlPjwvcmVjb3JkPjwvQ2l0ZT48L0VuZE5vdGU+

ADDIN EN.CITE.DATA 2, 3, 6Supplementary Figure S4. Cytotransmabs internalizes into various types of live cells with internalizing efficiency of almost 100%. (A) Cellular internalization and localization of cytotransmabs (TMab4, HuT4 and AvaT4) and control mAbs (adalimumab and bevacizumab) in PANC-1, HT-29 and MCF-7 cells, treated with the antibodies (1 ?M) for 6 h at 37 °C. Image magnification, 630× (zoom factor 2); scale bar, 5 μm. (B) Internalization efficiency of cytotransmabs (TMab4, HuT4 and AvaT4) observed for numerous cells of HeLa, PANC-1, HT-29 and MCF-7, treated with the antibodies (1 ?M) for 6 h at 37 °C. The images are representative of hundreds of cells examined by confocal microscopy. Image magnification, 400× (zoom factor 1); scale bar, 20 μm. In (A and B), after removing cell-surface bound antibodies with low pH glycine buffer (pH 2.5), internalized antibodies were visualized with FITC-conjugated anti-human IgG Fc antibody. Images show the merging of antibodies (green) and Hochest33342-stained nuclei (blue) at the centered single confocal section. (C) Representative FACS scatter plots of HeLa cells, treated with the indicated antibodies (1 ?M) for 6 h at 37 °C prior to FACS analysis, showing the internalization efficiency of cytotransmabs (TMab4 and AvaT4) and the control mAb (Bevacizumab). Internalized antibodies were stained with Alexa Fluor 488-conjugated anti-human IgG antibody. 'Control' means the cells left untreated with antibodies. Gating (rectangles) indicate the cells with Alexa Fluor 488 positive fluorescence compared to the untreated control. The indicated percentages were the frequency of positive cell population entering the boxed gate.Supplementary Figure S5. Cytotoxicity of internalized cytotransmabs in PANC-1 cells, treated with the indicated antibodies (1 ?M) at 37 °C for 24 h or 48 h. (A) The morphological features of cells were taken by phase-contrast microscopy after 48 h. Image magnification, 100×; scale bar, 20 μm. (B) The cell viability was determined by MTT assay after 24 h or 48 h. The data represent mean ± SD versus PBS-treated controls.Supplementary Figure S6. Confocal fluorescence images showing effects of endocytosis inhibitors on TMab4 internalization into HeLa cells, as described in Fig. 3E. HeLa cells were pre-treated for 30 min at 37 °C with CPZ (10 μg/ml), dansylcadaverine (200 μM), MβCD (5 mM), nystatin (50 μg/ml), EIPA (10 μM), wortmannin (200 nM), Cyt-D (1μg/ml) or dynasore (40 μM) and then incubated with TMab4 for additional 2 h. Internalized TMab4 was stained with TRITC-conjugated anti-human IgG Fc antibody and analyzed by confocal microscopy. Images show the merging of TMab4 (red) and Hochest33342-stained nuclei (blue) at the centered single confocal section. Scale bar represents 5 m.Supplementary Figure S7. Time-lapse live cell imaging to monitor cellular internalization and endosomal release of TMab4. HeLa cells were incubated with Alexa Flour 488-labeled TMab4 (1 μM) and real-time internalization of TMab4 was observed by time-lapse imaging. The dead time between TMab4 treatment and image taking was about 5 min. (A and B) Montage image (A) and quantification of TMab4 internalization and cytosolic accumulation (B), monitored for 2 h with 3 min interval. In (A), representative montage images showing cellular internalization and localization, taken at the indicated times at the centered single confocal section during 2 h TMab4 internalization. Left-top DIC image was added to show cytosolic region and nucleus of cells. Cell boundary was drawn with green line and nucleus was filled with blue color. Image magnification, 630×; scale bar, 10 μm. In (B), measurement of TMab4 fluorescence intensity in the cytosolic regions (n = 13 cells) with the incubation time. Data are presented as mean ± SEM. Supplementary Figure S8. Kinetic parameters for the interactions of C20 and KT4 IgG antibodies with soluble KRS, monitored by SPR. The binding affinity of the anti-KRS antibodies were measured using purified protein by surface resonance plasmon method measured using a BIAcore 2000. (A) SPR sensograms showing the kinetic interactions of C20 and KT4 with surface-immobilized KRS. (B) Kinetic interaction parameters obtained from (A) data using BIAevaluation software. aThe equilibrium dissociation constants (KD1) were calculated from the relationship of KD1 = kd1 / ka1 using the bivalent binding model. bThe apparent equilibrium dissociation constants (KD) including the avidity binding were analyzed using the 1:1 Langmuir model. Each value represents the mean ± SD of two independent experiments. In each experiment, at least five data sets were used in the determination of the kinetic constants.Supplementary Movie S1. Live cell imaging to monitor cellular internalization and endosomal release of Alexa488-labeled TMab4. Movies showing the internalization and endosomal escape of TMab4, tracked for 2 h with 3 min interval. Rapid internalization of TMab4 was indicated by the appearance of small intracellular punctuate structures, the number of which increased with time in the cytoplasmic regions. The cytosolic release of TMab4 was indicated by gradual increase diffuse fluorescence signal intensity in the cytosolic regions.Supplementary Materials and MethodsReagents and antibodies Chlorpromazine (CPZ, C8138), dansylcadaverine (30432), methyl-β-cyclodextrin (MβCD, C4555), nystatin (N6261), 5-(N-Ethyl-N-isopropyl) amiloride (EIPA, A3085), wortmannin (W1628), cytochalasin D (Cyt-D, C8273), dynasore (D7693), MG132 (M7449), rabbit IgG (I5006), FITC-cholera toxin B (Ctx-B, C1655), fluorescein isothiocyanate (FITC) conjugated anti-human IgG (Fc specific, F9512) and tetramethyl-rhodamine isothiocyanate (TRITC) conjugated anti-rabbit IgG (T6778) antibodies were purchased from Sigma-Aldrich. Alexa 488-transferrin (TF, T13342), Oregon green-dextran (D7173), LysoTracker? Red DND-99 (L7528) and were obtained from Invitrogen (Molecular Probes). Horse radish peroxidase (HRP) conjugated anti-mouse IgG (Fc specific, #7076) and anti-p53 (#9282) antibodies were from Cell Signaling. HRP conjugated anti-rabbit IgG (Fc specific, sc-2004), HRP-conjugated anti-goat IgG (Fc specific, sc-2033), anti-β-actin (sc-69879), anti-EEA1 (sc-53939), anti-caveolin-1 (sc-53564), anti-calnexin (sc-70481) and anti-58K Golgi protein (sc-58770) antibodies were purchased from Santa Cruz Biotechnology. Anti-human IgG (Fc specific, 31125) antibody was from Thermo scientific. TRITC-conjugated anti-human IgG (Fc specific, H581) antibody was from Leinco Technologies and anti-KRS mAb was from BIOCON (Medicinal Bioconvergence Research Center, Gyeonggi, Korea).PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5LaW08L0F1dGhvcj48WWVhcj4yMDE0PC9ZZWFyPjxSZWNO

dW0+MzQ4PC9SZWNOdW0+PERpc3BsYXlUZXh0PjxzdHlsZSBmYWNlPSJzdXBlcnNjcmlwdCI+Nzwv

c3R5bGU+PC9EaXNwbGF5VGV4dD48cmVjb3JkPjxyZWMtbnVtYmVyPjM0ODwvcmVjLW51bWJlcj48

Zm9yZWlnbi1rZXlzPjxrZXkgYXBwPSJFTiIgZGItaWQ9IjVzejBlNTJ6c3Nhd3hjZWEyeHBwdzU1

OGZyejV6enMyZDAydiI+MzQ4PC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10eXBlIG5hbWU9Ikpv

dXJuYWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1dGhvcnM+PGF1dGhv

cj5LaW0sIEQuIEcuPC9hdXRob3I+PGF1dGhvcj5MZWUsIEouIFkuPC9hdXRob3I+PGF1dGhvcj5L

d29uLCBOLiBILjwvYXV0aG9yPjxhdXRob3I+RmFuZywgUC48L2F1dGhvcj48YXV0aG9yPlpoYW5n

LCBRLjwvYXV0aG9yPjxhdXRob3I+V2FuZywgSi48L2F1dGhvcj48YXV0aG9yPllvdW5nLCBOLiBM

LjwvYXV0aG9yPjxhdXRob3I+R3VvLCBNLjwvYXV0aG9yPjxhdXRob3I+Q2hvLCBILiBZLjwvYXV0

aG9yPjxhdXRob3I+TXVzaHRhcSwgQS4gVS48L2F1dGhvcj48YXV0aG9yPkplb24sIFkuIEguPC9h

dXRob3I+PGF1dGhvcj5DaG9pLCBKLiBXLjwvYXV0aG9yPjxhdXRob3I+SGFuLCBKLiBNLjwvYXV0

aG9yPjxhdXRob3I+S2FuZywgSC4gVy48L2F1dGhvcj48YXV0aG9yPkpvbywgSi4gRS48L2F1dGhv

cj48YXV0aG9yPkh1ciwgWS48L2F1dGhvcj48YXV0aG9yPkthbmcsIFcuPC9hdXRob3I+PGF1dGhv

cj5ZYW5nLCBILjwvYXV0aG9yPjxhdXRob3I+TmFtLCBELiBILjwvYXV0aG9yPjxhdXRob3I+TGVl

LCBNLiBTLjwvYXV0aG9yPjxhdXRob3I+TGVlLCBKLiBXLjwvYXV0aG9yPjxhdXRob3I+S2ltLCBF

LiBTLjwvYXV0aG9yPjxhdXRob3I+TW9vbiwgQS48L2F1dGhvcj48YXV0aG9yPktpbSwgSy48L2F1

dGhvcj48YXV0aG9yPktpbSwgRC48L2F1dGhvcj48YXV0aG9yPkthbmcsIEUuIEouPC9hdXRob3I+

PGF1dGhvcj5Nb29uLCBZLjwvYXV0aG9yPjxhdXRob3I+UmhlZSwgSy4gSC48L2F1dGhvcj48YXV0

aG9yPkhhbiwgQi4gVy48L2F1dGhvcj48YXV0aG9yPllhbmcsIEouIFMuPC9hdXRob3I+PGF1dGhv

cj5IYW4sIEcuPC9hdXRob3I+PGF1dGhvcj5ZYW5nLCBXLiBTLjwvYXV0aG9yPjxhdXRob3I+TGVl

LCBDLjwvYXV0aG9yPjxhdXRob3I+V2FuZywgTS4gVy48L2F1dGhvcj48YXV0aG9yPktpbSwgUy48

L2F1dGhvcj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVzcz4xXSBNZWRpY2lu

YWwgQmlvY29udmVyZ2VuY2UgUmVzZWFyY2ggQ2VudGVyLCBTZW91bCBOYXRpb25hbCBVbml2ZXJz

aXR5LCBTZW91bCwgS29yZWEuIFsyXSBSZXNlYXJjaCBJbnN0aXR1dGUgb2YgUGhhcm1hY2V1dGlj

YWwgU2NpZW5jZXMsIENvbGxlZ2Ugb2YgUGhhcm1hY3ksIFNlb3VsIE5hdGlvbmFsIFVuaXZlcnNp

dHksIFNlb3VsLCBLb3JlYS4gWzNdLiYjeEQ7MV0gTWVkaWNpbmFsIEJpb2NvbnZlcmdlbmNlIFJl

c2VhcmNoIENlbnRlciwgU2VvdWwgTmF0aW9uYWwgVW5pdmVyc2l0eSwgU2VvdWwsIEtvcmVhLiBb

Ml0gUmVzZWFyY2ggSW5zdGl0dXRlIG9mIFBoYXJtYWNldXRpY2FsIFNjaWVuY2VzLCBDb2xsZWdl

IG9mIFBoYXJtYWN5LCBTZW91bCBOYXRpb25hbCBVbml2ZXJzaXR5LCBTZW91bCwgS29yZWEuJiN4

RDtEZXBhcnRtZW50IG9mIENhbmNlciBCaW9sb2d5LCBUaGUgU2NyaXBwcyBSZXNlYXJjaCBJbnN0

aXR1dGUsIFNjcmlwcHMgRmxvcmlkYSwgSnVwaXRlciwgRmxvcmlkYSwgVVNBLiYjeEQ7RGVwYXJ0

bWVudCBvZiBDaGVtaXN0cnkgYW5kIEJpb2NoZW1pc3RyeSwgRmxvcmlkYSBTdGF0ZSBVbml2ZXJz

aXR5LCBUYWxsYWhhc3NlZSwgRmxvcmlkYSwgVVNBLiYjeEQ7SW9uIEN5Y2xvdHJvbiBSZXNvbmFu

Y2UgUHJvZ3JhbSwgTmF0aW9uYWwgSGlnaCBNYWduZXRpYyBGaWVsZCBMYWJvcmF0b3J5LCBGbG9y

aWRhIFN0YXRlIFVuaXZlcnNpdHksIFRhbGxhaGFzc2VlLCBGbG9yaWRhLCBVU0EuJiN4RDtDb2xs

ZWdlIG9mIFBoYXJtYWN5LCBLb3JlYSBVbml2ZXJzaXR5LCBTZWpvbmcsIEtvcmVhLiYjeEQ7MV0g

TWVkaWNpbmFsIEJpb2NvbnZlcmdlbmNlIFJlc2VhcmNoIENlbnRlciwgU2VvdWwgTmF0aW9uYWwg

VW5pdmVyc2l0eSwgU2VvdWwsIEtvcmVhLiBbMl0gV2VsbG1hbiBDZW50ZXIgZm9yIFBob3RvbWVk

aWNpbmUsIE1hc3NhY2h1c2V0dHMgR2VuZXJhbCBIb3NwaXRhbCwgSGFydmFyZCBNZWRpY2FsIFNj

aG9vbCwgQm9zdG9uLCBNYXNzYWNodXNldHRzLCBVU0EuJiN4RDtZdWhhbiBSZXNlYXJjaCBJbnN0

aXR1dGUsIFlvbmdpbiwgS29yZWEuJiN4RDtEZXBhcnRtZW50IG9mIE5ldXJvc3VyZ2VyeSwgU2Ft

c3VuZyBNZWRpY2FsIENlbnRlciwgU3VuZ2t5dW5rd2FuIFVuaXZlcnNpdHkgU2Nob29sIG9mIE1l

ZGljaW5lLCBTZW91bCwgS29yZWEuJiN4RDtSZXNlYXJjaCBJbnN0aXR1dGUgb2YgUGhhcm1hY2V1

dGljYWwgU2NpZW5jZXMsIENvbGxlZ2Ugb2YgUGhhcm1hY3ksIFNlb3VsIE5hdGlvbmFsIFVuaXZl

cnNpdHksIFNlb3VsLCBLb3JlYS4mI3hEO0NvbGxlZ2Ugb2YgUGhhcm1hY3ksIER1a3N1bmcgV29t

ZW4mYXBvcztzIFVuaXZlcnNpdHksIFNlb3VsLCBLb3JlYS4mI3hEO01lZGljaW5hbCBCaW9jb252

ZXJnZW5jZSBSZXNlYXJjaCBDZW50ZXIsIFNlb3VsIE5hdGlvbmFsIFVuaXZlcnNpdHksIFNlb3Vs

LCBLb3JlYS4mI3hEO1RyYW5zbGF0aW9uYWwgUmVzZWFyY2ggQ2VudGVyIGZvciBQcm90ZWluIEZ1

bmN0aW9uIENvbnRyb2wsIERlcGFydG1lbnQgb2YgQmlvdGVjaG5vbG9neSBhbmQgV0NVIERlcGFy

dG1lbnQgb2YgQmlvbWVkaWNhbCBTY2llbmNlcywgWW9uc2VpIFVuaXZlcnNpdHksIFNlb3VsLCBL

b3JlYS4mI3hEO0JSSSwgS29yZWEgSW5zdGl0dXRlIG9mIFNjaWVuY2UgYW5kIFRlY2hub2xvZ3ks

IFNlb3VsLCBLb3JlYS4mI3hEO1RoZSBOYXRpb25hbCBDZW50ZXIgZm9yIERydWcgU2NyZWVuaW5n

LCBaaGFuZ2ppYW5nIEhpZ2gtVGVjaCBQYXJrLCBTaGFuZ2hhaSwgQ2hpbmEuJiN4RDsxXSBNZWRp

Y2luYWwgQmlvY29udmVyZ2VuY2UgUmVzZWFyY2ggQ2VudGVyLCBTZW91bCBOYXRpb25hbCBVbml2

ZXJzaXR5LCBTZW91bCwgS29yZWEuIFsyXSBSZXNlYXJjaCBJbnN0aXR1dGUgb2YgUGhhcm1hY2V1

dGljYWwgU2NpZW5jZXMsIENvbGxlZ2Ugb2YgUGhhcm1hY3ksIFNlb3VsIE5hdGlvbmFsIFVuaXZl

cnNpdHksIFNlb3VsLCBLb3JlYS4gWzNdIFdvcmxkIENsYXNzIFVuaXZlcnNpdHkgRGVwYXJ0bWVu

dCBvZiBNb2xlY3VsYXIgTWVkaWNpbmUgYW5kIEJpb3BoYXJtYWNldXRpY2FsIFNjaWVuY2VzLCBT

ZW91bCBOYXRpb25hbCBVbml2ZXJzaXR5LCBTZW91bCwgS29yZWEuPC9hdXRoLWFkZHJlc3M+PHRp

dGxlcz48dGl0bGU+Q2hlbWljYWwgaW5oaWJpdGlvbiBvZiBwcm9tZXRhc3RhdGljIGx5c3lsLXRS

TkEgc3ludGhldGFzZS1sYW1pbmluIHJlY2VwdG9yIGludGVyYWN0aW9uPC90aXRsZT48c2Vjb25k

YXJ5LXRpdGxlPk5hdCBDaGVtIEJpb2w8L3NlY29uZGFyeS10aXRsZT48YWx0LXRpdGxlPk5hdHVy

ZSBjaGVtaWNhbCBiaW9sb2d5PC9hbHQtdGl0bGU+PC90aXRsZXM+PHBlcmlvZGljYWw+PGZ1bGwt

dGl0bGU+TmF0IENoZW0gQmlvbDwvZnVsbC10aXRsZT48YWJici0xPk5hdHVyZSBjaGVtaWNhbCBi

aW9sb2d5PC9hYmJyLTE+PC9wZXJpb2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVsbC10aXRsZT5O

YXQgQ2hlbSBCaW9sPC9mdWxsLXRpdGxlPjxhYmJyLTE+TmF0dXJlIGNoZW1pY2FsIGJpb2xvZ3k8

L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4yOS0zNDwvcGFnZXM+PHZvbHVtZT4xMDwv

dm9sdW1lPjxudW1iZXI+MTwvbnVtYmVyPjxrZXl3b3Jkcz48a2V5d29yZD5DZWxsIE1lbWJyYW5l

L21ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+THlzaW5lLXRSTkEgTGlnYXNlL2FudGFnb25p

c3RzICZhbXA7IGluaGliaXRvcnMvKm1ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+Kk5lb3Bs

YXNtIE1ldGFzdGFzaXM8L2tleXdvcmQ+PGtleXdvcmQ+UHJvdGVpbiBUcmFuc3BvcnQ8L2tleXdv

cmQ+PGtleXdvcmQ+UmVjZXB0b3JzLCBMYW1pbmluL2FudGFnb25pc3RzICZhbXA7IGluaGliaXRv

cnMvKm1ldGFib2xpc208L2tleXdvcmQ+PC9rZXl3b3Jkcz48ZGF0ZXM+PHllYXI+MjAxNDwveWVh

cj48cHViLWRhdGVzPjxkYXRlPkphbjwvZGF0ZT48L3B1Yi1kYXRlcz48L2RhdGVzPjxpc2JuPjE1

NTItNDQ2OSAoRWxlY3Ryb25pYykmI3hEOzE1NTItNDQ1MCAoTGlua2luZyk8L2lzYm4+PGFjY2Vz

c2lvbi1udW0+MjQyMTIxMzY8L2FjY2Vzc2lvbi1udW0+PHVybHM+PHJlbGF0ZWQtdXJscz48dXJs

Pmh0dHA6Ly93d3cubmNiaS5ubG0ubmloLmdvdi9wdWJtZWQvMjQyMTIxMzY8L3VybD48L3JlbGF0

ZWQtdXJscz48L3VybHM+PGVsZWN0cm9uaWMtcmVzb3VyY2UtbnVtPjEwLjEwMzgvbmNoZW1iaW8u

MTM4MTwvZWxlY3Ryb25pYy1yZXNvdXJjZS1udW0+PC9yZWNvcmQ+PC9DaXRlPjwvRW5kTm90ZT4A

ADDIN EN.CITE PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5LaW08L0F1dGhvcj48WWVhcj4yMDE0PC9ZZWFyPjxSZWNO

dW0+MzQ4PC9SZWNOdW0+PERpc3BsYXlUZXh0PjxzdHlsZSBmYWNlPSJzdXBlcnNjcmlwdCI+Nzwv

c3R5bGU+PC9EaXNwbGF5VGV4dD48cmVjb3JkPjxyZWMtbnVtYmVyPjM0ODwvcmVjLW51bWJlcj48

Zm9yZWlnbi1rZXlzPjxrZXkgYXBwPSJFTiIgZGItaWQ9IjVzejBlNTJ6c3Nhd3hjZWEyeHBwdzU1

OGZyejV6enMyZDAydiI+MzQ4PC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10eXBlIG5hbWU9Ikpv

dXJuYWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1dGhvcnM+PGF1dGhv

cj5LaW0sIEQuIEcuPC9hdXRob3I+PGF1dGhvcj5MZWUsIEouIFkuPC9hdXRob3I+PGF1dGhvcj5L

d29uLCBOLiBILjwvYXV0aG9yPjxhdXRob3I+RmFuZywgUC48L2F1dGhvcj48YXV0aG9yPlpoYW5n

LCBRLjwvYXV0aG9yPjxhdXRob3I+V2FuZywgSi48L2F1dGhvcj48YXV0aG9yPllvdW5nLCBOLiBM

LjwvYXV0aG9yPjxhdXRob3I+R3VvLCBNLjwvYXV0aG9yPjxhdXRob3I+Q2hvLCBILiBZLjwvYXV0

aG9yPjxhdXRob3I+TXVzaHRhcSwgQS4gVS48L2F1dGhvcj48YXV0aG9yPkplb24sIFkuIEguPC9h

dXRob3I+PGF1dGhvcj5DaG9pLCBKLiBXLjwvYXV0aG9yPjxhdXRob3I+SGFuLCBKLiBNLjwvYXV0

aG9yPjxhdXRob3I+S2FuZywgSC4gVy48L2F1dGhvcj48YXV0aG9yPkpvbywgSi4gRS48L2F1dGhv

cj48YXV0aG9yPkh1ciwgWS48L2F1dGhvcj48YXV0aG9yPkthbmcsIFcuPC9hdXRob3I+PGF1dGhv

cj5ZYW5nLCBILjwvYXV0aG9yPjxhdXRob3I+TmFtLCBELiBILjwvYXV0aG9yPjxhdXRob3I+TGVl

LCBNLiBTLjwvYXV0aG9yPjxhdXRob3I+TGVlLCBKLiBXLjwvYXV0aG9yPjxhdXRob3I+S2ltLCBF

LiBTLjwvYXV0aG9yPjxhdXRob3I+TW9vbiwgQS48L2F1dGhvcj48YXV0aG9yPktpbSwgSy48L2F1

dGhvcj48YXV0aG9yPktpbSwgRC48L2F1dGhvcj48YXV0aG9yPkthbmcsIEUuIEouPC9hdXRob3I+

PGF1dGhvcj5Nb29uLCBZLjwvYXV0aG9yPjxhdXRob3I+UmhlZSwgSy4gSC48L2F1dGhvcj48YXV0

aG9yPkhhbiwgQi4gVy48L2F1dGhvcj48YXV0aG9yPllhbmcsIEouIFMuPC9hdXRob3I+PGF1dGhv

cj5IYW4sIEcuPC9hdXRob3I+PGF1dGhvcj5ZYW5nLCBXLiBTLjwvYXV0aG9yPjxhdXRob3I+TGVl

LCBDLjwvYXV0aG9yPjxhdXRob3I+V2FuZywgTS4gVy48L2F1dGhvcj48YXV0aG9yPktpbSwgUy48

L2F1dGhvcj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVzcz4xXSBNZWRpY2lu

YWwgQmlvY29udmVyZ2VuY2UgUmVzZWFyY2ggQ2VudGVyLCBTZW91bCBOYXRpb25hbCBVbml2ZXJz

aXR5LCBTZW91bCwgS29yZWEuIFsyXSBSZXNlYXJjaCBJbnN0aXR1dGUgb2YgUGhhcm1hY2V1dGlj

YWwgU2NpZW5jZXMsIENvbGxlZ2Ugb2YgUGhhcm1hY3ksIFNlb3VsIE5hdGlvbmFsIFVuaXZlcnNp

dHksIFNlb3VsLCBLb3JlYS4gWzNdLiYjeEQ7MV0gTWVkaWNpbmFsIEJpb2NvbnZlcmdlbmNlIFJl

c2VhcmNoIENlbnRlciwgU2VvdWwgTmF0aW9uYWwgVW5pdmVyc2l0eSwgU2VvdWwsIEtvcmVhLiBb

Ml0gUmVzZWFyY2ggSW5zdGl0dXRlIG9mIFBoYXJtYWNldXRpY2FsIFNjaWVuY2VzLCBDb2xsZWdl

IG9mIFBoYXJtYWN5LCBTZW91bCBOYXRpb25hbCBVbml2ZXJzaXR5LCBTZW91bCwgS29yZWEuJiN4

RDtEZXBhcnRtZW50IG9mIENhbmNlciBCaW9sb2d5LCBUaGUgU2NyaXBwcyBSZXNlYXJjaCBJbnN0

aXR1dGUsIFNjcmlwcHMgRmxvcmlkYSwgSnVwaXRlciwgRmxvcmlkYSwgVVNBLiYjeEQ7RGVwYXJ0

bWVudCBvZiBDaGVtaXN0cnkgYW5kIEJpb2NoZW1pc3RyeSwgRmxvcmlkYSBTdGF0ZSBVbml2ZXJz

aXR5LCBUYWxsYWhhc3NlZSwgRmxvcmlkYSwgVVNBLiYjeEQ7SW9uIEN5Y2xvdHJvbiBSZXNvbmFu

Y2UgUHJvZ3JhbSwgTmF0aW9uYWwgSGlnaCBNYWduZXRpYyBGaWVsZCBMYWJvcmF0b3J5LCBGbG9y

aWRhIFN0YXRlIFVuaXZlcnNpdHksIFRhbGxhaGFzc2VlLCBGbG9yaWRhLCBVU0EuJiN4RDtDb2xs

ZWdlIG9mIFBoYXJtYWN5LCBLb3JlYSBVbml2ZXJzaXR5LCBTZWpvbmcsIEtvcmVhLiYjeEQ7MV0g

TWVkaWNpbmFsIEJpb2NvbnZlcmdlbmNlIFJlc2VhcmNoIENlbnRlciwgU2VvdWwgTmF0aW9uYWwg

VW5pdmVyc2l0eSwgU2VvdWwsIEtvcmVhLiBbMl0gV2VsbG1hbiBDZW50ZXIgZm9yIFBob3RvbWVk

aWNpbmUsIE1hc3NhY2h1c2V0dHMgR2VuZXJhbCBIb3NwaXRhbCwgSGFydmFyZCBNZWRpY2FsIFNj

aG9vbCwgQm9zdG9uLCBNYXNzYWNodXNldHRzLCBVU0EuJiN4RDtZdWhhbiBSZXNlYXJjaCBJbnN0

aXR1dGUsIFlvbmdpbiwgS29yZWEuJiN4RDtEZXBhcnRtZW50IG9mIE5ldXJvc3VyZ2VyeSwgU2Ft

c3VuZyBNZWRpY2FsIENlbnRlciwgU3VuZ2t5dW5rd2FuIFVuaXZlcnNpdHkgU2Nob29sIG9mIE1l

ZGljaW5lLCBTZW91bCwgS29yZWEuJiN4RDtSZXNlYXJjaCBJbnN0aXR1dGUgb2YgUGhhcm1hY2V1

dGljYWwgU2NpZW5jZXMsIENvbGxlZ2Ugb2YgUGhhcm1hY3ksIFNlb3VsIE5hdGlvbmFsIFVuaXZl

cnNpdHksIFNlb3VsLCBLb3JlYS4mI3hEO0NvbGxlZ2Ugb2YgUGhhcm1hY3ksIER1a3N1bmcgV29t

ZW4mYXBvcztzIFVuaXZlcnNpdHksIFNlb3VsLCBLb3JlYS4mI3hEO01lZGljaW5hbCBCaW9jb252

ZXJnZW5jZSBSZXNlYXJjaCBDZW50ZXIsIFNlb3VsIE5hdGlvbmFsIFVuaXZlcnNpdHksIFNlb3Vs

LCBLb3JlYS4mI3hEO1RyYW5zbGF0aW9uYWwgUmVzZWFyY2ggQ2VudGVyIGZvciBQcm90ZWluIEZ1

bmN0aW9uIENvbnRyb2wsIERlcGFydG1lbnQgb2YgQmlvdGVjaG5vbG9neSBhbmQgV0NVIERlcGFy

dG1lbnQgb2YgQmlvbWVkaWNhbCBTY2llbmNlcywgWW9uc2VpIFVuaXZlcnNpdHksIFNlb3VsLCBL

b3JlYS4mI3hEO0JSSSwgS29yZWEgSW5zdGl0dXRlIG9mIFNjaWVuY2UgYW5kIFRlY2hub2xvZ3ks

IFNlb3VsLCBLb3JlYS4mI3hEO1RoZSBOYXRpb25hbCBDZW50ZXIgZm9yIERydWcgU2NyZWVuaW5n

LCBaaGFuZ2ppYW5nIEhpZ2gtVGVjaCBQYXJrLCBTaGFuZ2hhaSwgQ2hpbmEuJiN4RDsxXSBNZWRp

Y2luYWwgQmlvY29udmVyZ2VuY2UgUmVzZWFyY2ggQ2VudGVyLCBTZW91bCBOYXRpb25hbCBVbml2

ZXJzaXR5LCBTZW91bCwgS29yZWEuIFsyXSBSZXNlYXJjaCBJbnN0aXR1dGUgb2YgUGhhcm1hY2V1

dGljYWwgU2NpZW5jZXMsIENvbGxlZ2Ugb2YgUGhhcm1hY3ksIFNlb3VsIE5hdGlvbmFsIFVuaXZl

cnNpdHksIFNlb3VsLCBLb3JlYS4gWzNdIFdvcmxkIENsYXNzIFVuaXZlcnNpdHkgRGVwYXJ0bWVu

dCBvZiBNb2xlY3VsYXIgTWVkaWNpbmUgYW5kIEJpb3BoYXJtYWNldXRpY2FsIFNjaWVuY2VzLCBT

ZW91bCBOYXRpb25hbCBVbml2ZXJzaXR5LCBTZW91bCwgS29yZWEuPC9hdXRoLWFkZHJlc3M+PHRp

dGxlcz48dGl0bGU+Q2hlbWljYWwgaW5oaWJpdGlvbiBvZiBwcm9tZXRhc3RhdGljIGx5c3lsLXRS

TkEgc3ludGhldGFzZS1sYW1pbmluIHJlY2VwdG9yIGludGVyYWN0aW9uPC90aXRsZT48c2Vjb25k

YXJ5LXRpdGxlPk5hdCBDaGVtIEJpb2w8L3NlY29uZGFyeS10aXRsZT48YWx0LXRpdGxlPk5hdHVy

ZSBjaGVtaWNhbCBiaW9sb2d5PC9hbHQtdGl0bGU+PC90aXRsZXM+PHBlcmlvZGljYWw+PGZ1bGwt

dGl0bGU+TmF0IENoZW0gQmlvbDwvZnVsbC10aXRsZT48YWJici0xPk5hdHVyZSBjaGVtaWNhbCBi

aW9sb2d5PC9hYmJyLTE+PC9wZXJpb2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVsbC10aXRsZT5O

YXQgQ2hlbSBCaW9sPC9mdWxsLXRpdGxlPjxhYmJyLTE+TmF0dXJlIGNoZW1pY2FsIGJpb2xvZ3k8

L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4yOS0zNDwvcGFnZXM+PHZvbHVtZT4xMDwv

dm9sdW1lPjxudW1iZXI+MTwvbnVtYmVyPjxrZXl3b3Jkcz48a2V5d29yZD5DZWxsIE1lbWJyYW5l

L21ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+THlzaW5lLXRSTkEgTGlnYXNlL2FudGFnb25p

c3RzICZhbXA7IGluaGliaXRvcnMvKm1ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+Kk5lb3Bs

YXNtIE1ldGFzdGFzaXM8L2tleXdvcmQ+PGtleXdvcmQ+UHJvdGVpbiBUcmFuc3BvcnQ8L2tleXdv

cmQ+PGtleXdvcmQ+UmVjZXB0b3JzLCBMYW1pbmluL2FudGFnb25pc3RzICZhbXA7IGluaGliaXRv

cnMvKm1ldGFib2xpc208L2tleXdvcmQ+PC9rZXl3b3Jkcz48ZGF0ZXM+PHllYXI+MjAxNDwveWVh

cj48cHViLWRhdGVzPjxkYXRlPkphbjwvZGF0ZT48L3B1Yi1kYXRlcz48L2RhdGVzPjxpc2JuPjE1

NTItNDQ2OSAoRWxlY3Ryb25pYykmI3hEOzE1NTItNDQ1MCAoTGlua2luZyk8L2lzYm4+PGFjY2Vz

c2lvbi1udW0+MjQyMTIxMzY8L2FjY2Vzc2lvbi1udW0+PHVybHM+PHJlbGF0ZWQtdXJscz48dXJs

Pmh0dHA6Ly93d3cubmNiaS5ubG0ubmloLmdvdi9wdWJtZWQvMjQyMTIxMzY8L3VybD48L3JlbGF0

ZWQtdXJscz48L3VybHM+PGVsZWN0cm9uaWMtcmVzb3VyY2UtbnVtPjEwLjEwMzgvbmNoZW1iaW8u

MTM4MTwvZWxlY3Ryb25pYy1yZXNvdXJjZS1udW0+PC9yZWNvcmQ+PC9DaXRlPjwvRW5kTm90ZT4A

ADDIN EN.CITE.DATA 7 The recombinant proteins, KRS, histidyl-tRNA synthetase (HRS), tryptophanyl-tRNA synthetase (WRS), and ARS-interacting multifunctional protein 1 (AIMP1), and were provided from BIOCON.PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5LaW08L0F1dGhvcj48WWVhcj4yMDE0PC9ZZWFyPjxSZWNO

dW0+MzQ4PC9SZWNOdW0+PERpc3BsYXlUZXh0PjxzdHlsZSBmYWNlPSJzdXBlcnNjcmlwdCI+Nzwv

c3R5bGU+PC9EaXNwbGF5VGV4dD48cmVjb3JkPjxyZWMtbnVtYmVyPjM0ODwvcmVjLW51bWJlcj48

Zm9yZWlnbi1rZXlzPjxrZXkgYXBwPSJFTiIgZGItaWQ9IjVzejBlNTJ6c3Nhd3hjZWEyeHBwdzU1

OGZyejV6enMyZDAydiI+MzQ4PC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10eXBlIG5hbWU9Ikpv

dXJuYWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1dGhvcnM+PGF1dGhv

cj5LaW0sIEQuIEcuPC9hdXRob3I+PGF1dGhvcj5MZWUsIEouIFkuPC9hdXRob3I+PGF1dGhvcj5L

d29uLCBOLiBILjwvYXV0aG9yPjxhdXRob3I+RmFuZywgUC48L2F1dGhvcj48YXV0aG9yPlpoYW5n

LCBRLjwvYXV0aG9yPjxhdXRob3I+V2FuZywgSi48L2F1dGhvcj48YXV0aG9yPllvdW5nLCBOLiBM

LjwvYXV0aG9yPjxhdXRob3I+R3VvLCBNLjwvYXV0aG9yPjxhdXRob3I+Q2hvLCBILiBZLjwvYXV0

aG9yPjxhdXRob3I+TXVzaHRhcSwgQS4gVS48L2F1dGhvcj48YXV0aG9yPkplb24sIFkuIEguPC9h

dXRob3I+PGF1dGhvcj5DaG9pLCBKLiBXLjwvYXV0aG9yPjxhdXRob3I+SGFuLCBKLiBNLjwvYXV0

aG9yPjxhdXRob3I+S2FuZywgSC4gVy48L2F1dGhvcj48YXV0aG9yPkpvbywgSi4gRS48L2F1dGhv

cj48YXV0aG9yPkh1ciwgWS48L2F1dGhvcj48YXV0aG9yPkthbmcsIFcuPC9hdXRob3I+PGF1dGhv

cj5ZYW5nLCBILjwvYXV0aG9yPjxhdXRob3I+TmFtLCBELiBILjwvYXV0aG9yPjxhdXRob3I+TGVl

LCBNLiBTLjwvYXV0aG9yPjxhdXRob3I+TGVlLCBKLiBXLjwvYXV0aG9yPjxhdXRob3I+S2ltLCBF

LiBTLjwvYXV0aG9yPjxhdXRob3I+TW9vbiwgQS48L2F1dGhvcj48YXV0aG9yPktpbSwgSy48L2F1

dGhvcj48YXV0aG9yPktpbSwgRC48L2F1dGhvcj48YXV0aG9yPkthbmcsIEUuIEouPC9hdXRob3I+

PGF1dGhvcj5Nb29uLCBZLjwvYXV0aG9yPjxhdXRob3I+UmhlZSwgSy4gSC48L2F1dGhvcj48YXV0

aG9yPkhhbiwgQi4gVy48L2F1dGhvcj48YXV0aG9yPllhbmcsIEouIFMuPC9hdXRob3I+PGF1dGhv

cj5IYW4sIEcuPC9hdXRob3I+PGF1dGhvcj5ZYW5nLCBXLiBTLjwvYXV0aG9yPjxhdXRob3I+TGVl

LCBDLjwvYXV0aG9yPjxhdXRob3I+V2FuZywgTS4gVy48L2F1dGhvcj48YXV0aG9yPktpbSwgUy48

L2F1dGhvcj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVzcz4xXSBNZWRpY2lu

YWwgQmlvY29udmVyZ2VuY2UgUmVzZWFyY2ggQ2VudGVyLCBTZW91bCBOYXRpb25hbCBVbml2ZXJz

aXR5LCBTZW91bCwgS29yZWEuIFsyXSBSZXNlYXJjaCBJbnN0aXR1dGUgb2YgUGhhcm1hY2V1dGlj

YWwgU2NpZW5jZXMsIENvbGxlZ2Ugb2YgUGhhcm1hY3ksIFNlb3VsIE5hdGlvbmFsIFVuaXZlcnNp

dHksIFNlb3VsLCBLb3JlYS4gWzNdLiYjeEQ7MV0gTWVkaWNpbmFsIEJpb2NvbnZlcmdlbmNlIFJl

c2VhcmNoIENlbnRlciwgU2VvdWwgTmF0aW9uYWwgVW5pdmVyc2l0eSwgU2VvdWwsIEtvcmVhLiBb

Ml0gUmVzZWFyY2ggSW5zdGl0dXRlIG9mIFBoYXJtYWNldXRpY2FsIFNjaWVuY2VzLCBDb2xsZWdl

IG9mIFBoYXJtYWN5LCBTZW91bCBOYXRpb25hbCBVbml2ZXJzaXR5LCBTZW91bCwgS29yZWEuJiN4

RDtEZXBhcnRtZW50IG9mIENhbmNlciBCaW9sb2d5LCBUaGUgU2NyaXBwcyBSZXNlYXJjaCBJbnN0

aXR1dGUsIFNjcmlwcHMgRmxvcmlkYSwgSnVwaXRlciwgRmxvcmlkYSwgVVNBLiYjeEQ7RGVwYXJ0

bWVudCBvZiBDaGVtaXN0cnkgYW5kIEJpb2NoZW1pc3RyeSwgRmxvcmlkYSBTdGF0ZSBVbml2ZXJz

aXR5LCBUYWxsYWhhc3NlZSwgRmxvcmlkYSwgVVNBLiYjeEQ7SW9uIEN5Y2xvdHJvbiBSZXNvbmFu

Y2UgUHJvZ3JhbSwgTmF0aW9uYWwgSGlnaCBNYWduZXRpYyBGaWVsZCBMYWJvcmF0b3J5LCBGbG9y

aWRhIFN0YXRlIFVuaXZlcnNpdHksIFRhbGxhaGFzc2VlLCBGbG9yaWRhLCBVU0EuJiN4RDtDb2xs

ZWdlIG9mIFBoYXJtYWN5LCBLb3JlYSBVbml2ZXJzaXR5LCBTZWpvbmcsIEtvcmVhLiYjeEQ7MV0g

TWVkaWNpbmFsIEJpb2NvbnZlcmdlbmNlIFJlc2VhcmNoIENlbnRlciwgU2VvdWwgTmF0aW9uYWwg

VW5pdmVyc2l0eSwgU2VvdWwsIEtvcmVhLiBbMl0gV2VsbG1hbiBDZW50ZXIgZm9yIFBob3RvbWVk

aWNpbmUsIE1hc3NhY2h1c2V0dHMgR2VuZXJhbCBIb3NwaXRhbCwgSGFydmFyZCBNZWRpY2FsIFNj

aG9vbCwgQm9zdG9uLCBNYXNzYWNodXNldHRzLCBVU0EuJiN4RDtZdWhhbiBSZXNlYXJjaCBJbnN0

aXR1dGUsIFlvbmdpbiwgS29yZWEuJiN4RDtEZXBhcnRtZW50IG9mIE5ldXJvc3VyZ2VyeSwgU2Ft

c3VuZyBNZWRpY2FsIENlbnRlciwgU3VuZ2t5dW5rd2FuIFVuaXZlcnNpdHkgU2Nob29sIG9mIE1l

ZGljaW5lLCBTZW91bCwgS29yZWEuJiN4RDtSZXNlYXJjaCBJbnN0aXR1dGUgb2YgUGhhcm1hY2V1

dGljYWwgU2NpZW5jZXMsIENvbGxlZ2Ugb2YgUGhhcm1hY3ksIFNlb3VsIE5hdGlvbmFsIFVuaXZl

cnNpdHksIFNlb3VsLCBLb3JlYS4mI3hEO0NvbGxlZ2Ugb2YgUGhhcm1hY3ksIER1a3N1bmcgV29t

ZW4mYXBvcztzIFVuaXZlcnNpdHksIFNlb3VsLCBLb3JlYS4mI3hEO01lZGljaW5hbCBCaW9jb252

ZXJnZW5jZSBSZXNlYXJjaCBDZW50ZXIsIFNlb3VsIE5hdGlvbmFsIFVuaXZlcnNpdHksIFNlb3Vs

LCBLb3JlYS4mI3hEO1RyYW5zbGF0aW9uYWwgUmVzZWFyY2ggQ2VudGVyIGZvciBQcm90ZWluIEZ1

bmN0aW9uIENvbnRyb2wsIERlcGFydG1lbnQgb2YgQmlvdGVjaG5vbG9neSBhbmQgV0NVIERlcGFy

dG1lbnQgb2YgQmlvbWVkaWNhbCBTY2llbmNlcywgWW9uc2VpIFVuaXZlcnNpdHksIFNlb3VsLCBL

b3JlYS4mI3hEO0JSSSwgS29yZWEgSW5zdGl0dXRlIG9mIFNjaWVuY2UgYW5kIFRlY2hub2xvZ3ks

IFNlb3VsLCBLb3JlYS4mI3hEO1RoZSBOYXRpb25hbCBDZW50ZXIgZm9yIERydWcgU2NyZWVuaW5n

LCBaaGFuZ2ppYW5nIEhpZ2gtVGVjaCBQYXJrLCBTaGFuZ2hhaSwgQ2hpbmEuJiN4RDsxXSBNZWRp

Y2luYWwgQmlvY29udmVyZ2VuY2UgUmVzZWFyY2ggQ2VudGVyLCBTZW91bCBOYXRpb25hbCBVbml2

ZXJzaXR5LCBTZW91bCwgS29yZWEuIFsyXSBSZXNlYXJjaCBJbnN0aXR1dGUgb2YgUGhhcm1hY2V1

dGljYWwgU2NpZW5jZXMsIENvbGxlZ2Ugb2YgUGhhcm1hY3ksIFNlb3VsIE5hdGlvbmFsIFVuaXZl

cnNpdHksIFNlb3VsLCBLb3JlYS4gWzNdIFdvcmxkIENsYXNzIFVuaXZlcnNpdHkgRGVwYXJ0bWVu

dCBvZiBNb2xlY3VsYXIgTWVkaWNpbmUgYW5kIEJpb3BoYXJtYWNldXRpY2FsIFNjaWVuY2VzLCBT

ZW91bCBOYXRpb25hbCBVbml2ZXJzaXR5LCBTZW91bCwgS29yZWEuPC9hdXRoLWFkZHJlc3M+PHRp

dGxlcz48dGl0bGU+Q2hlbWljYWwgaW5oaWJpdGlvbiBvZiBwcm9tZXRhc3RhdGljIGx5c3lsLXRS

TkEgc3ludGhldGFzZS1sYW1pbmluIHJlY2VwdG9yIGludGVyYWN0aW9uPC90aXRsZT48c2Vjb25k

YXJ5LXRpdGxlPk5hdCBDaGVtIEJpb2w8L3NlY29uZGFyeS10aXRsZT48YWx0LXRpdGxlPk5hdHVy

ZSBjaGVtaWNhbCBiaW9sb2d5PC9hbHQtdGl0bGU+PC90aXRsZXM+PHBlcmlvZGljYWw+PGZ1bGwt

dGl0bGU+TmF0IENoZW0gQmlvbDwvZnVsbC10aXRsZT48YWJici0xPk5hdHVyZSBjaGVtaWNhbCBi

aW9sb2d5PC9hYmJyLTE+PC9wZXJpb2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVsbC10aXRsZT5O

YXQgQ2hlbSBCaW9sPC9mdWxsLXRpdGxlPjxhYmJyLTE+TmF0dXJlIGNoZW1pY2FsIGJpb2xvZ3k8

L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4yOS0zNDwvcGFnZXM+PHZvbHVtZT4xMDwv

dm9sdW1lPjxudW1iZXI+MTwvbnVtYmVyPjxrZXl3b3Jkcz48a2V5d29yZD5DZWxsIE1lbWJyYW5l

L21ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+THlzaW5lLXRSTkEgTGlnYXNlL2FudGFnb25p

c3RzICZhbXA7IGluaGliaXRvcnMvKm1ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+Kk5lb3Bs

YXNtIE1ldGFzdGFzaXM8L2tleXdvcmQ+PGtleXdvcmQ+UHJvdGVpbiBUcmFuc3BvcnQ8L2tleXdv

cmQ+PGtleXdvcmQ+UmVjZXB0b3JzLCBMYW1pbmluL2FudGFnb25pc3RzICZhbXA7IGluaGliaXRv

cnMvKm1ldGFib2xpc208L2tleXdvcmQ+PC9rZXl3b3Jkcz48ZGF0ZXM+PHllYXI+MjAxNDwveWVh

cj48cHViLWRhdGVzPjxkYXRlPkphbjwvZGF0ZT48L3B1Yi1kYXRlcz48L2RhdGVzPjxpc2JuPjE1

NTItNDQ2OSAoRWxlY3Ryb25pYykmI3hEOzE1NTItNDQ1MCAoTGlua2luZyk8L2lzYm4+PGFjY2Vz

c2lvbi1udW0+MjQyMTIxMzY8L2FjY2Vzc2lvbi1udW0+PHVybHM+PHJlbGF0ZWQtdXJscz48dXJs

Pmh0dHA6Ly93d3cubmNiaS5ubG0ubmloLmdvdi9wdWJtZWQvMjQyMTIxMzY8L3VybD48L3JlbGF0

ZWQtdXJscz48L3VybHM+PGVsZWN0cm9uaWMtcmVzb3VyY2UtbnVtPjEwLjEwMzgvbmNoZW1iaW8u

MTM4MTwvZWxlY3Ryb25pYy1yZXNvdXJjZS1udW0+PC9yZWNvcmQ+PC9DaXRlPjwvRW5kTm90ZT4A

ADDIN EN.CITE PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5LaW08L0F1dGhvcj48WWVhcj4yMDE0PC9ZZWFyPjxSZWNO

dW0+MzQ4PC9SZWNOdW0+PERpc3BsYXlUZXh0PjxzdHlsZSBmYWNlPSJzdXBlcnNjcmlwdCI+Nzwv

c3R5bGU+PC9EaXNwbGF5VGV4dD48cmVjb3JkPjxyZWMtbnVtYmVyPjM0ODwvcmVjLW51bWJlcj48

Zm9yZWlnbi1rZXlzPjxrZXkgYXBwPSJFTiIgZGItaWQ9IjVzejBlNTJ6c3Nhd3hjZWEyeHBwdzU1

OGZyejV6enMyZDAydiI+MzQ4PC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10eXBlIG5hbWU9Ikpv

dXJuYWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1dGhvcnM+PGF1dGhv

cj5LaW0sIEQuIEcuPC9hdXRob3I+PGF1dGhvcj5MZWUsIEouIFkuPC9hdXRob3I+PGF1dGhvcj5L

d29uLCBOLiBILjwvYXV0aG9yPjxhdXRob3I+RmFuZywgUC48L2F1dGhvcj48YXV0aG9yPlpoYW5n

LCBRLjwvYXV0aG9yPjxhdXRob3I+V2FuZywgSi48L2F1dGhvcj48YXV0aG9yPllvdW5nLCBOLiBM

LjwvYXV0aG9yPjxhdXRob3I+R3VvLCBNLjwvYXV0aG9yPjxhdXRob3I+Q2hvLCBILiBZLjwvYXV0

aG9yPjxhdXRob3I+TXVzaHRhcSwgQS4gVS48L2F1dGhvcj48YXV0aG9yPkplb24sIFkuIEguPC9h

dXRob3I+PGF1dGhvcj5DaG9pLCBKLiBXLjwvYXV0aG9yPjxhdXRob3I+SGFuLCBKLiBNLjwvYXV0

aG9yPjxhdXRob3I+S2FuZywgSC4gVy48L2F1dGhvcj48YXV0aG9yPkpvbywgSi4gRS48L2F1dGhv

cj48YXV0aG9yPkh1ciwgWS48L2F1dGhvcj48YXV0aG9yPkthbmcsIFcuPC9hdXRob3I+PGF1dGhv

cj5ZYW5nLCBILjwvYXV0aG9yPjxhdXRob3I+TmFtLCBELiBILjwvYXV0aG9yPjxhdXRob3I+TGVl

LCBNLiBTLjwvYXV0aG9yPjxhdXRob3I+TGVlLCBKLiBXLjwvYXV0aG9yPjxhdXRob3I+S2ltLCBF

LiBTLjwvYXV0aG9yPjxhdXRob3I+TW9vbiwgQS48L2F1dGhvcj48YXV0aG9yPktpbSwgSy48L2F1

dGhvcj48YXV0aG9yPktpbSwgRC48L2F1dGhvcj48YXV0aG9yPkthbmcsIEUuIEouPC9hdXRob3I+

PGF1dGhvcj5Nb29uLCBZLjwvYXV0aG9yPjxhdXRob3I+UmhlZSwgSy4gSC48L2F1dGhvcj48YXV0

aG9yPkhhbiwgQi4gVy48L2F1dGhvcj48YXV0aG9yPllhbmcsIEouIFMuPC9hdXRob3I+PGF1dGhv

cj5IYW4sIEcuPC9hdXRob3I+PGF1dGhvcj5ZYW5nLCBXLiBTLjwvYXV0aG9yPjxhdXRob3I+TGVl

LCBDLjwvYXV0aG9yPjxhdXRob3I+V2FuZywgTS4gVy48L2F1dGhvcj48YXV0aG9yPktpbSwgUy48

L2F1dGhvcj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVzcz4xXSBNZWRpY2lu

YWwgQmlvY29udmVyZ2VuY2UgUmVzZWFyY2ggQ2VudGVyLCBTZW91bCBOYXRpb25hbCBVbml2ZXJz

aXR5LCBTZW91bCwgS29yZWEuIFsyXSBSZXNlYXJjaCBJbnN0aXR1dGUgb2YgUGhhcm1hY2V1dGlj

YWwgU2NpZW5jZXMsIENvbGxlZ2Ugb2YgUGhhcm1hY3ksIFNlb3VsIE5hdGlvbmFsIFVuaXZlcnNp

dHksIFNlb3VsLCBLb3JlYS4gWzNdLiYjeEQ7MV0gTWVkaWNpbmFsIEJpb2NvbnZlcmdlbmNlIFJl

c2VhcmNoIENlbnRlciwgU2VvdWwgTmF0aW9uYWwgVW5pdmVyc2l0eSwgU2VvdWwsIEtvcmVhLiBb

Ml0gUmVzZWFyY2ggSW5zdGl0dXRlIG9mIFBoYXJtYWNldXRpY2FsIFNjaWVuY2VzLCBDb2xsZWdl

IG9mIFBoYXJtYWN5LCBTZW91bCBOYXRpb25hbCBVbml2ZXJzaXR5LCBTZW91bCwgS29yZWEuJiN4

RDtEZXBhcnRtZW50IG9mIENhbmNlciBCaW9sb2d5LCBUaGUgU2NyaXBwcyBSZXNlYXJjaCBJbnN0

aXR1dGUsIFNjcmlwcHMgRmxvcmlkYSwgSnVwaXRlciwgRmxvcmlkYSwgVVNBLiYjeEQ7RGVwYXJ0

bWVudCBvZiBDaGVtaXN0cnkgYW5kIEJpb2NoZW1pc3RyeSwgRmxvcmlkYSBTdGF0ZSBVbml2ZXJz

aXR5LCBUYWxsYWhhc3NlZSwgRmxvcmlkYSwgVVNBLiYjeEQ7SW9uIEN5Y2xvdHJvbiBSZXNvbmFu

Y2UgUHJvZ3JhbSwgTmF0aW9uYWwgSGlnaCBNYWduZXRpYyBGaWVsZCBMYWJvcmF0b3J5LCBGbG9y

aWRhIFN0YXRlIFVuaXZlcnNpdHksIFRhbGxhaGFzc2VlLCBGbG9yaWRhLCBVU0EuJiN4RDtDb2xs

ZWdlIG9mIFBoYXJtYWN5LCBLb3JlYSBVbml2ZXJzaXR5LCBTZWpvbmcsIEtvcmVhLiYjeEQ7MV0g

TWVkaWNpbmFsIEJpb2NvbnZlcmdlbmNlIFJlc2VhcmNoIENlbnRlciwgU2VvdWwgTmF0aW9uYWwg

VW5pdmVyc2l0eSwgU2VvdWwsIEtvcmVhLiBbMl0gV2VsbG1hbiBDZW50ZXIgZm9yIFBob3RvbWVk

aWNpbmUsIE1hc3NhY2h1c2V0dHMgR2VuZXJhbCBIb3NwaXRhbCwgSGFydmFyZCBNZWRpY2FsIFNj

aG9vbCwgQm9zdG9uLCBNYXNzYWNodXNldHRzLCBVU0EuJiN4RDtZdWhhbiBSZXNlYXJjaCBJbnN0

aXR1dGUsIFlvbmdpbiwgS29yZWEuJiN4RDtEZXBhcnRtZW50IG9mIE5ldXJvc3VyZ2VyeSwgU2Ft

c3VuZyBNZWRpY2FsIENlbnRlciwgU3VuZ2t5dW5rd2FuIFVuaXZlcnNpdHkgU2Nob29sIG9mIE1l

ZGljaW5lLCBTZW91bCwgS29yZWEuJiN4RDtSZXNlYXJjaCBJbnN0aXR1dGUgb2YgUGhhcm1hY2V1

dGljYWwgU2NpZW5jZXMsIENvbGxlZ2Ugb2YgUGhhcm1hY3ksIFNlb3VsIE5hdGlvbmFsIFVuaXZl

cnNpdHksIFNlb3VsLCBLb3JlYS4mI3hEO0NvbGxlZ2Ugb2YgUGhhcm1hY3ksIER1a3N1bmcgV29t

ZW4mYXBvcztzIFVuaXZlcnNpdHksIFNlb3VsLCBLb3JlYS4mI3hEO01lZGljaW5hbCBCaW9jb252

ZXJnZW5jZSBSZXNlYXJjaCBDZW50ZXIsIFNlb3VsIE5hdGlvbmFsIFVuaXZlcnNpdHksIFNlb3Vs

LCBLb3JlYS4mI3hEO1RyYW5zbGF0aW9uYWwgUmVzZWFyY2ggQ2VudGVyIGZvciBQcm90ZWluIEZ1

bmN0aW9uIENvbnRyb2wsIERlcGFydG1lbnQgb2YgQmlvdGVjaG5vbG9neSBhbmQgV0NVIERlcGFy

dG1lbnQgb2YgQmlvbWVkaWNhbCBTY2llbmNlcywgWW9uc2VpIFVuaXZlcnNpdHksIFNlb3VsLCBL

b3JlYS4mI3hEO0JSSSwgS29yZWEgSW5zdGl0dXRlIG9mIFNjaWVuY2UgYW5kIFRlY2hub2xvZ3ks

IFNlb3VsLCBLb3JlYS4mI3hEO1RoZSBOYXRpb25hbCBDZW50ZXIgZm9yIERydWcgU2NyZWVuaW5n

LCBaaGFuZ2ppYW5nIEhpZ2gtVGVjaCBQYXJrLCBTaGFuZ2hhaSwgQ2hpbmEuJiN4RDsxXSBNZWRp

Y2luYWwgQmlvY29udmVyZ2VuY2UgUmVzZWFyY2ggQ2VudGVyLCBTZW91bCBOYXRpb25hbCBVbml2

ZXJzaXR5LCBTZW91bCwgS29yZWEuIFsyXSBSZXNlYXJjaCBJbnN0aXR1dGUgb2YgUGhhcm1hY2V1

dGljYWwgU2NpZW5jZXMsIENvbGxlZ2Ugb2YgUGhhcm1hY3ksIFNlb3VsIE5hdGlvbmFsIFVuaXZl

cnNpdHksIFNlb3VsLCBLb3JlYS4gWzNdIFdvcmxkIENsYXNzIFVuaXZlcnNpdHkgRGVwYXJ0bWVu

dCBvZiBNb2xlY3VsYXIgTWVkaWNpbmUgYW5kIEJpb3BoYXJtYWNldXRpY2FsIFNjaWVuY2VzLCBT

ZW91bCBOYXRpb25hbCBVbml2ZXJzaXR5LCBTZW91bCwgS29yZWEuPC9hdXRoLWFkZHJlc3M+PHRp

dGxlcz48dGl0bGU+Q2hlbWljYWwgaW5oaWJpdGlvbiBvZiBwcm9tZXRhc3RhdGljIGx5c3lsLXRS

TkEgc3ludGhldGFzZS1sYW1pbmluIHJlY2VwdG9yIGludGVyYWN0aW9uPC90aXRsZT48c2Vjb25k

YXJ5LXRpdGxlPk5hdCBDaGVtIEJpb2w8L3NlY29uZGFyeS10aXRsZT48YWx0LXRpdGxlPk5hdHVy

ZSBjaGVtaWNhbCBiaW9sb2d5PC9hbHQtdGl0bGU+PC90aXRsZXM+PHBlcmlvZGljYWw+PGZ1bGwt

dGl0bGU+TmF0IENoZW0gQmlvbDwvZnVsbC10aXRsZT48YWJici0xPk5hdHVyZSBjaGVtaWNhbCBi

aW9sb2d5PC9hYmJyLTE+PC9wZXJpb2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVsbC10aXRsZT5O

YXQgQ2hlbSBCaW9sPC9mdWxsLXRpdGxlPjxhYmJyLTE+TmF0dXJlIGNoZW1pY2FsIGJpb2xvZ3k8

L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4yOS0zNDwvcGFnZXM+PHZvbHVtZT4xMDwv

dm9sdW1lPjxudW1iZXI+MTwvbnVtYmVyPjxrZXl3b3Jkcz48a2V5d29yZD5DZWxsIE1lbWJyYW5l

L21ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+THlzaW5lLXRSTkEgTGlnYXNlL2FudGFnb25p

c3RzICZhbXA7IGluaGliaXRvcnMvKm1ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+Kk5lb3Bs

YXNtIE1ldGFzdGFzaXM8L2tleXdvcmQ+PGtleXdvcmQ+UHJvdGVpbiBUcmFuc3BvcnQ8L2tleXdv

cmQ+PGtleXdvcmQ+UmVjZXB0b3JzLCBMYW1pbmluL2FudGFnb25pc3RzICZhbXA7IGluaGliaXRv

cnMvKm1ldGFib2xpc208L2tleXdvcmQ+PC9rZXl3b3Jkcz48ZGF0ZXM+PHllYXI+MjAxNDwveWVh

cj48cHViLWRhdGVzPjxkYXRlPkphbjwvZGF0ZT48L3B1Yi1kYXRlcz48L2RhdGVzPjxpc2JuPjE1

NTItNDQ2OSAoRWxlY3Ryb25pYykmI3hEOzE1NTItNDQ1MCAoTGlua2luZyk8L2lzYm4+PGFjY2Vz

c2lvbi1udW0+MjQyMTIxMzY8L2FjY2Vzc2lvbi1udW0+PHVybHM+PHJlbGF0ZWQtdXJscz48dXJs

Pmh0dHA6Ly93d3cubmNiaS5ubG0ubmloLmdvdi9wdWJtZWQvMjQyMTIxMzY8L3VybD48L3JlbGF0

ZWQtdXJscz48L3VybHM+PGVsZWN0cm9uaWMtcmVzb3VyY2UtbnVtPjEwLjEwMzgvbmNoZW1iaW8u

MTM4MTwvZWxlY3Ryb25pYy1yZXNvdXJjZS1udW0+PC9yZWNvcmQ+PC9DaXRlPjwvRW5kTm90ZT4A

ADDIN EN.CITE.DATA 7Size Exclusion Chromatography (SEC) SEC analyses were performed on the Agilent 1100 high performance liquid chromatography system using a superdexTM200 10/300GC (10 mm × 300 mm, GE Healthcare) size-exclusion column PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5MZWU8L0F1dGhvcj48WWVhcj4yMDEwPC9ZZWFyPjxSZWNO

dW0+NTc8L1JlY051bT48RGlzcGxheVRleHQ+PHN0eWxlIGZhY2U9InN1cGVyc2NyaXB0Ij44PC9z

dHlsZT48L0Rpc3BsYXlUZXh0PjxyZWNvcmQ+PHJlYy1udW1iZXI+NTc8L3JlYy1udW1iZXI+PGZv

cmVpZ24ta2V5cz48a2V5IGFwcD0iRU4iIGRiLWlkPSI1c3owZTUyenNzYXd4Y2VhMnhwcHc1NThm

cno1enpzMmQwMnYiPjU3PC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10eXBlIG5hbWU9IkpvdXJu

YWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1dGhvcnM+PGF1dGhvcj5M

ZWUsIFMuIEguPC9hdXRob3I+PGF1dGhvcj5QYXJrLCBELiBXLjwvYXV0aG9yPjxhdXRob3I+U3Vu

ZywgRS4gUy48L2F1dGhvcj48YXV0aG9yPlBhcmssIEguIFIuPC9hdXRob3I+PGF1dGhvcj5LaW0s

IEouIEsuPC9hdXRob3I+PGF1dGhvcj5LaW0sIFkuIFMuPC9hdXRob3I+PC9hdXRob3JzPjwvY29u

dHJpYnV0b3JzPjxhdXRoLWFkZHJlc3M+RGVwdC4gb2YgTW9sZWN1bGFyIFNjaWVuY2UgYW5kIFRl

Y2hub2xvZ3ksIEFqb3UgVW5pdmVyc2l0eSwgU3V3b24gNDQzLTc0OSwgUmVwdWJsaWMgb2YgS29y

ZWEuPC9hdXRoLWFkZHJlc3M+PHRpdGxlcz48dGl0bGU+SHVtYW5pemF0aW9uIG9mIGFuIGFnb25p

c3RpYyBhbnRpLWRlYXRoIHJlY2VwdG9yIDQgc2luZ2xlIGNoYWluIHZhcmlhYmxlIGZyYWdtZW50

IGFudGlib2R5IGFuZCBhdmlkaXR5LW1lZGlhdGVkIGVuaGFuY2VtZW50IG9mIGl0cyBjZWxsIGRl

YXRoLWluZHVjaW5nIGFjdGl2aXR5PC90aXRsZT48c2Vjb25kYXJ5LXRpdGxlPk1vbCBJbW11bm9s

PC9zZWNvbmRhcnktdGl0bGU+PC90aXRsZXM+PHBlcmlvZGljYWw+PGZ1bGwtdGl0bGU+TW9sIElt

bXVub2w8L2Z1bGwtdGl0bGU+PC9wZXJpb2RpY2FsPjxwYWdlcz44MTYtMjQ8L3BhZ2VzPjx2b2x1

bWU+NDc8L3ZvbHVtZT48bnVtYmVyPjQ8L251bWJlcj48a2V5d29yZHM+PGtleXdvcmQ+QW1pbm8g

QWNpZCBTZXF1ZW5jZTwva2V5d29yZD48a2V5d29yZD5BbmltYWxzPC9rZXl3b3JkPjxrZXl3b3Jk

PkFudGlib2RpZXMvY2hlbWlzdHJ5LyppbW11bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPkFudGli

b2R5IEFmZmluaXR5LyppbW11bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPipDeXRvdG94aWNpdHks

IEltbXVub2xvZ2ljPC9rZXl3b3JkPjxrZXl3b3JkPkhDVDExNiBDZWxsczwva2V5d29yZD48a2V5

d29yZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdvcmQ+SW1tb2JpbGl6ZWQgUHJvdGVpbnMvY2hlbWlz

dHJ5L2ltbXVub2xvZ3k8L2tleXdvcmQ+PGtleXdvcmQ+SW1tdW5vZ2xvYnVsaW4gRnJhZ21lbnRz

L2NoZW1pc3RyeS8qaW1tdW5vbG9neTwva2V5d29yZD48a2V5d29yZD5JbW11bm9nbG9idWxpbiBW

YXJpYWJsZSBSZWdpb24vY2hlbWlzdHJ5LyppbW11bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPktp

bmV0aWNzPC9rZXl3b3JkPjxrZXl3b3JkPk1pY2U8L2tleXdvcmQ+PGtleXdvcmQ+TW9sZWN1bGFy

IFNlcXVlbmNlIERhdGE8L2tleXdvcmQ+PGtleXdvcmQ+UHJvdGVpbiBTdHJ1Y3R1cmUsIFRlcnRp

YXJ5PC9rZXl3b3JkPjxrZXl3b3JkPlJlY2VwdG9ycywgVE5GLVJlbGF0ZWQgQXBvcHRvc2lzLUlu

ZHVjaW5nPC9rZXl3b3JkPjxrZXl3b3JkPkxpZ2FuZC8qYWdvbmlzdHMvY2hlbWlzdHJ5LyppbW11

bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPlJlY29tYmluYW50IEZ1c2lvbiBQcm90ZWlucy9jaGVt

aXN0cnkvaW1tdW5vbG9neTwva2V5d29yZD48a2V5d29yZD5TZXF1ZW5jZSBBbGlnbm1lbnQ8L2tl

eXdvcmQ+PC9rZXl3b3Jkcz48ZGF0ZXM+PHllYXI+MjAxMDwveWVhcj48cHViLWRhdGVzPjxkYXRl

PkphbjwvZGF0ZT48L3B1Yi1kYXRlcz48L2RhdGVzPjxhY2Nlc3Npb24tbnVtPjE5ODY0MDI3PC9h

Y2Nlc3Npb24tbnVtPjx1cmxzPjxyZWxhdGVkLXVybHM+PHVybD5odHRwOi8vd3d3Lm5jYmkubmxt

Lm5paC5nb3YvZW50cmV6L3F1ZXJ5LmZjZ2k/Y21kPVJldHJpZXZlJmFtcDtkYj1QdWJNZWQmYW1w

O2RvcHQ9Q2l0YXRpb24mYW1wO2xpc3RfdWlkcz0xOTg2NDAyNyA8L3VybD48L3JlbGF0ZWQtdXJs

cz48L3VybHM+PC9yZWNvcmQ+PC9DaXRlPjwvRW5kTm90ZT5=

ADDIN EN.CITE PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5MZWU8L0F1dGhvcj48WWVhcj4yMDEwPC9ZZWFyPjxSZWNO

dW0+NTc8L1JlY051bT48RGlzcGxheVRleHQ+PHN0eWxlIGZhY2U9InN1cGVyc2NyaXB0Ij44PC9z

dHlsZT48L0Rpc3BsYXlUZXh0PjxyZWNvcmQ+PHJlYy1udW1iZXI+NTc8L3JlYy1udW1iZXI+PGZv

cmVpZ24ta2V5cz48a2V5IGFwcD0iRU4iIGRiLWlkPSI1c3owZTUyenNzYXd4Y2VhMnhwcHc1NThm

cno1enpzMmQwMnYiPjU3PC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10eXBlIG5hbWU9IkpvdXJu

YWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1dGhvcnM+PGF1dGhvcj5M

ZWUsIFMuIEguPC9hdXRob3I+PGF1dGhvcj5QYXJrLCBELiBXLjwvYXV0aG9yPjxhdXRob3I+U3Vu

ZywgRS4gUy48L2F1dGhvcj48YXV0aG9yPlBhcmssIEguIFIuPC9hdXRob3I+PGF1dGhvcj5LaW0s

IEouIEsuPC9hdXRob3I+PGF1dGhvcj5LaW0sIFkuIFMuPC9hdXRob3I+PC9hdXRob3JzPjwvY29u

dHJpYnV0b3JzPjxhdXRoLWFkZHJlc3M+RGVwdC4gb2YgTW9sZWN1bGFyIFNjaWVuY2UgYW5kIFRl

Y2hub2xvZ3ksIEFqb3UgVW5pdmVyc2l0eSwgU3V3b24gNDQzLTc0OSwgUmVwdWJsaWMgb2YgS29y

ZWEuPC9hdXRoLWFkZHJlc3M+PHRpdGxlcz48dGl0bGU+SHVtYW5pemF0aW9uIG9mIGFuIGFnb25p

c3RpYyBhbnRpLWRlYXRoIHJlY2VwdG9yIDQgc2luZ2xlIGNoYWluIHZhcmlhYmxlIGZyYWdtZW50

IGFudGlib2R5IGFuZCBhdmlkaXR5LW1lZGlhdGVkIGVuaGFuY2VtZW50IG9mIGl0cyBjZWxsIGRl

YXRoLWluZHVjaW5nIGFjdGl2aXR5PC90aXRsZT48c2Vjb25kYXJ5LXRpdGxlPk1vbCBJbW11bm9s

PC9zZWNvbmRhcnktdGl0bGU+PC90aXRsZXM+PHBlcmlvZGljYWw+PGZ1bGwtdGl0bGU+TW9sIElt

bXVub2w8L2Z1bGwtdGl0bGU+PC9wZXJpb2RpY2FsPjxwYWdlcz44MTYtMjQ8L3BhZ2VzPjx2b2x1

bWU+NDc8L3ZvbHVtZT48bnVtYmVyPjQ8L251bWJlcj48a2V5d29yZHM+PGtleXdvcmQ+QW1pbm8g

QWNpZCBTZXF1ZW5jZTwva2V5d29yZD48a2V5d29yZD5BbmltYWxzPC9rZXl3b3JkPjxrZXl3b3Jk

PkFudGlib2RpZXMvY2hlbWlzdHJ5LyppbW11bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPkFudGli

b2R5IEFmZmluaXR5LyppbW11bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPipDeXRvdG94aWNpdHks

IEltbXVub2xvZ2ljPC9rZXl3b3JkPjxrZXl3b3JkPkhDVDExNiBDZWxsczwva2V5d29yZD48a2V5

d29yZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdvcmQ+SW1tb2JpbGl6ZWQgUHJvdGVpbnMvY2hlbWlz

dHJ5L2ltbXVub2xvZ3k8L2tleXdvcmQ+PGtleXdvcmQ+SW1tdW5vZ2xvYnVsaW4gRnJhZ21lbnRz

L2NoZW1pc3RyeS8qaW1tdW5vbG9neTwva2V5d29yZD48a2V5d29yZD5JbW11bm9nbG9idWxpbiBW

YXJpYWJsZSBSZWdpb24vY2hlbWlzdHJ5LyppbW11bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPktp

bmV0aWNzPC9rZXl3b3JkPjxrZXl3b3JkPk1pY2U8L2tleXdvcmQ+PGtleXdvcmQ+TW9sZWN1bGFy

IFNlcXVlbmNlIERhdGE8L2tleXdvcmQ+PGtleXdvcmQ+UHJvdGVpbiBTdHJ1Y3R1cmUsIFRlcnRp

YXJ5PC9rZXl3b3JkPjxrZXl3b3JkPlJlY2VwdG9ycywgVE5GLVJlbGF0ZWQgQXBvcHRvc2lzLUlu

ZHVjaW5nPC9rZXl3b3JkPjxrZXl3b3JkPkxpZ2FuZC8qYWdvbmlzdHMvY2hlbWlzdHJ5LyppbW11

bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPlJlY29tYmluYW50IEZ1c2lvbiBQcm90ZWlucy9jaGVt

aXN0cnkvaW1tdW5vbG9neTwva2V5d29yZD48a2V5d29yZD5TZXF1ZW5jZSBBbGlnbm1lbnQ8L2tl

eXdvcmQ+PC9rZXl3b3Jkcz48ZGF0ZXM+PHllYXI+MjAxMDwveWVhcj48cHViLWRhdGVzPjxkYXRl

PkphbjwvZGF0ZT48L3B1Yi1kYXRlcz48L2RhdGVzPjxhY2Nlc3Npb24tbnVtPjE5ODY0MDI3PC9h

Y2Nlc3Npb24tbnVtPjx1cmxzPjxyZWxhdGVkLXVybHM+PHVybD5odHRwOi8vd3d3Lm5jYmkubmxt

Lm5paC5nb3YvZW50cmV6L3F1ZXJ5LmZjZ2k/Y21kPVJldHJpZXZlJmFtcDtkYj1QdWJNZWQmYW1w

O2RvcHQ9Q2l0YXRpb24mYW1wO2xpc3RfdWlkcz0xOTg2NDAyNyA8L3VybD48L3JlbGF0ZWQtdXJs

cz48L3VybHM+PC9yZWNvcmQ+PC9DaXRlPjwvRW5kTm90ZT5=

ADDIN EN.CITE.DATA 8 equilibrated with 12 mM sodium phosphate buffer (pH 7.4) containing 500 mM NaCl, 2.7 mM KCl at a flow rate of 0.5 ml/min. Antibodies were analyzed by SEC at a concentration of 200 ?g/ml in 70 ?l of sample volume. Chromatograms were obtained by monitoring absorbance at 280 nm. The molecular mass of antibodies was estimated by calculating elution time into the calibration curve, obtained using standard molecular mass marker (alcohol dehydrogenase, 150 KDa; bovine serum albumin, 66 KDa; carbonic anhydrase, 29 KDa) (Sigma-Aldrich).ELISABinding specificity of antibodies to antigens was determined by ELISA. ELISA plates (Nunc, Invitrogen) were coated for 1 h at 4 °C with the indicated antigens (0.05 ?g/ml of VEGF, 0.1 ?g/ml of TNFα, or 1 ?g/ml of KRS, HRS, WRS, or AIMP1) and blocked with 5% (wt/vol) skim milk. After washing with PBST (PBS pH 7.4, 0.01% Tween-20), diluted antibodies in 5 % (wt/vol) skim milk were applied to each well for 1 h at 25 °C. After washing with PBST, bound antibodies were detected by labeling with alkaline phosphatase-conjugated goat anti-human IgG (Fc-specific) mAb (Sigma-Aldrich) and then incubating with p-nitrophenylphosphate. Absorbance was read at 405 nm on a VersaMax microplate reader (Molecular devices).Surface plasmon resonance (SPR)Kinetic interactions of antibody-protein were measured at 25 °C using Biacore 2000 (GE Healthcare), as previously described. ADDIN EN.CITE <EndNote><Cite><Author>Shin</Author><Year>2014</Year><RecNum>367</RecNum><DisplayText><style face="superscript">9</style></DisplayText><record><rec-number>367</rec-number><foreign-keys><key app="EN" db-id="5sz0e52zssawxcea2xppw558frz5zzs2d02v">367</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Shin, T. H.</author><author>Sung, E. S.</author><author>Kim, Y. J.</author><author>Kim, K. S.</author><author>Kim, S. H.</author><author>Kim, S. K.</author><author>Lee, Y. D.</author><author>Kim, Y. S.</author></authors></contributors><auth-address>Corresponding Author: Yong-Sung Kim, Ajou University, 206 Worldcup-ro, Suwon, 443-749, Korea. kimys@ajou.ac.kr.</auth-address><titles><title>Enhancement of the tumor penetration of monoclonal antibody by fusion of a neuropilin-targeting Peptide improves the antitumor efficacy</title><secondary-title>Mol Cancer Ther</secondary-title><alt-title>Molecular cancer therapeutics</alt-title></titles><periodical><full-title>Mol Cancer Ther</full-title><abbr-1>Molecular cancer therapeutics</abbr-1></periodical><alt-periodical><full-title>Mol Cancer Ther</full-title><abbr-1>Molecular cancer therapeutics</abbr-1></alt-periodical><pages>651-61</pages><volume>13</volume><number>3</number><dates><year>2014</year><pub-dates><date>Mar</date></pub-dates></dates><isbn>1538-8514 (Electronic)&#xD;1535-7163 (Linking)</isbn><accession-num>24435448</accession-num><urls><related-urls><url> After immobilization of KRS protein onto the carboxymethylated dextran surface of a CM5 sensor chip at a level of about 145 response units (RUs), various concentrations of antibodies including C20 (1.25 nM ~ 20 nM) and KT4 (6.25 nM ~ 100 nM) in PBS (pH 7.4) were injected into the flow cell at a flow rate of 30 ?l/min for 3 min with 3 min dissociation per cycles. After each cycle, surfaces were regenerated with a buffer (20 mM NaOH, 1 M NaCl, pH 10.0) for 1 min. The binding data were normalized by subtracting the response of a blank cell. Sensogram data were analyzed using BIAevaluation software version 3.1. The kinetic rate constants (ka1, kd1, ka2, and kd2), as well as equilibrium dissociation constant (KD1 = kd1 / ka1)), were estimated by global fitting analysis of the sensograms to the bivalent binding model.PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5MZWU8L0F1dGhvcj48WWVhcj4yMDEzPC9ZZWFyPjxSZWNO

dW0+NDQ5PC9SZWNOdW0+PERpc3BsYXlUZXh0PjxzdHlsZSBmYWNlPSJzdXBlcnNjcmlwdCI+MTAs

IDExPC9zdHlsZT48L0Rpc3BsYXlUZXh0PjxyZWNvcmQ+PHJlYy1udW1iZXI+NDQ5PC9yZWMtbnVt

YmVyPjxmb3JlaWduLWtleXM+PGtleSBhcHA9IkVOIiBkYi1pZD0iNXN6MGU1Mnpzc2F3eGNlYTJ4

cHB3NTU4ZnJ6NXp6czJkMDJ2Ij40NDk8L2tleT48L2ZvcmVpZ24ta2V5cz48cmVmLXR5cGUgbmFt

ZT0iSm91cm5hbCBBcnRpY2xlIj4xNzwvcmVmLXR5cGU+PGNvbnRyaWJ1dG9ycz48YXV0aG9ycz48

YXV0aG9yPkxlZSwgWS4gSC48L2F1dGhvcj48YXV0aG9yPklpamltYSwgTS48L2F1dGhvcj48YXV0

aG9yPkthZG8sIFkuPC9hdXRob3I+PGF1dGhvcj5NaXpvaGF0YSwgRS48L2F1dGhvcj48YXV0aG9y

Pklub3VlLCBULjwvYXV0aG9yPjxhdXRob3I+U3VnaXlhbWEsIEEuPC9hdXRob3I+PGF1dGhvcj5E

b2ksIEguPC9hdXRob3I+PGF1dGhvcj5TaGliYXNha2ksIFkuPC9hdXRob3I+PGF1dGhvcj5Lb2Rh

bWEsIFQuPC9hdXRob3I+PC9hdXRob3JzPjwvY29udHJpYnV0b3JzPjxhdXRoLWFkZHJlc3M+TGFi

b3JhdG9yeSBmb3IgU3lzdGVtcyBCaW9sb2d5IGFuZCBNZWRpY2luZSwgUmVzZWFyY2ggQ2VudGVy

IGZvciBBZHZhbmNlZCBTY2llbmNlIGFuZCBUZWNobm9sb2d5LCBUaGUgVW5pdmVyc2l0eSBvZiBU

b2t5bywgNC02LTEgS29tYWJhLCBNZWd1cm8ta3UsIFRva3lvIDE1My04OTA0LCBKYXBhbjsgRGVw

YXJ0bWVudCBvZiBMaWZlIFNjaWVuY2VzLCBHcmFkdWF0ZSBTY2hvb2wgb2YgQXJ0cyBhbmQgU2Np

ZW5jZXMsIFRoZSBVbml2ZXJzaXR5IG9mIFRva3lvLCAzLTgtMSBLb21hYmEsIE1lZ3Vyby1rdSwg

VG9reW8gMTUzLTg5MDIsIEphcGFuLjwvYXV0aC1hZGRyZXNzPjx0aXRsZXM+PHRpdGxlPkNvbnN0

cnVjdGlvbiBhbmQgY2hhcmFjdGVyaXphdGlvbiBvZiBmdW5jdGlvbmFsIGFudGktZXBpcmVndWxp

biBodW1hbml6ZWQgbW9ub2Nsb25hbCBhbnRpYm9kaWVzPC90aXRsZT48c2Vjb25kYXJ5LXRpdGxl

PkJpb2NoZW0gQmlvcGh5cyBSZXMgQ29tbXVuPC9zZWNvbmRhcnktdGl0bGU+PGFsdC10aXRsZT5C

aW9jaGVtaWNhbCBhbmQgYmlvcGh5c2ljYWwgcmVzZWFyY2ggY29tbXVuaWNhdGlvbnM8L2FsdC10

aXRsZT48L3RpdGxlcz48cGVyaW9kaWNhbD48ZnVsbC10aXRsZT5CaW9jaGVtIEJpb3BoeXMgUmVz

IENvbW11bjwvZnVsbC10aXRsZT48YWJici0xPkJpb2NoZW1pY2FsIGFuZCBiaW9waHlzaWNhbCBy

ZXNlYXJjaCBjb21tdW5pY2F0aW9uczwvYWJici0xPjwvcGVyaW9kaWNhbD48YWx0LXBlcmlvZGlj

YWw+PGZ1bGwtdGl0bGU+QmlvY2hlbSBCaW9waHlzIFJlcyBDb21tdW48L2Z1bGwtdGl0bGU+PGFi

YnItMT5CaW9jaGVtaWNhbCBhbmQgYmlvcGh5c2ljYWwgcmVzZWFyY2ggY29tbXVuaWNhdGlvbnM8

L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4xMDExLTc8L3BhZ2VzPjx2b2x1bWU+NDQx

PC92b2x1bWU+PG51bWJlcj40PC9udW1iZXI+PGtleXdvcmRzPjxrZXl3b3JkPkFtaW5vIEFjaWQg

U2VxdWVuY2U8L2tleXdvcmQ+PGtleXdvcmQ+QW50aWJvZGllcywgTW9ub2Nsb25hbCwgSHVtYW5p

emVkLypiaW9zeW50aGVzaXMvY2hlbWlzdHJ5L2ltbXVub2xvZ3k8L2tleXdvcmQ+PGtleXdvcmQ+

RXBpZGVybWFsIEdyb3d0aCBGYWN0b3IvYW50YWdvbmlzdHMgJmFtcDsgaW5oaWJpdG9ycy8qaW1t

dW5vbG9neTwva2V5d29yZD48a2V5d29yZD5IQ1QxMTYgQ2VsbHM8L2tleXdvcmQ+PGtleXdvcmQ+

SHVtYW5zPC9rZXl3b3JkPjxrZXl3b3JkPkltbXVub2dsb2J1bGluIFZhcmlhYmxlIFJlZ2lvbi9i

aW9zeW50aGVzaXMvY2hlbWlzdHJ5L2ltbXVub2xvZ3k8L2tleXdvcmQ+PGtleXdvcmQ+SnVya2F0

IENlbGxzPC9rZXl3b3JkPjxrZXl3b3JkPkx1Y2lmZXJhc2VzL2Jpb3N5bnRoZXNpczwva2V5d29y

ZD48a2V5d29yZD5Nb2RlbHMsIE1vbGVjdWxhcjwva2V5d29yZD48a2V5d29yZD5Nb2xlY3VsYXIg

U2VxdWVuY2UgRGF0YTwva2V5d29yZD48a2V5d29yZD5ORkFUQyBUcmFuc2NyaXB0aW9uIEZhY3Rv

cnMvbWV0YWJvbGlzbTwva2V5d29yZD48a2V5d29yZD5Qcm90ZWluIEVuZ2luZWVyaW5nPC9rZXl3

b3JkPjxrZXl3b3JkPlJlY2VwdG9ycywgSWdHL2Jpb3N5bnRoZXNpczwva2V5d29yZD48L2tleXdv

cmRzPjxkYXRlcz48eWVhcj4yMDEzPC95ZWFyPjxwdWItZGF0ZXM+PGRhdGU+Tm92IDI5PC9kYXRl

PjwvcHViLWRhdGVzPjwvZGF0ZXM+PGlzYm4+MTA5MC0yMTA0IChFbGVjdHJvbmljKSYjeEQ7MDAw

Ni0yOTFYIChMaW5raW5nKTwvaXNibj48YWNjZXNzaW9uLW51bT4yNDIzOTU0OTwvYWNjZXNzaW9u

LW51bT48dXJscz48cmVsYXRlZC11cmxzPjx1cmw+aHR0cDovL3d3dy5uY2JpLm5sbS5uaWguZ292

L3B1Ym1lZC8yNDIzOTU0OTwvdXJsPjwvcmVsYXRlZC11cmxzPjwvdXJscz48ZWxlY3Ryb25pYy1y

ZXNvdXJjZS1udW0+MTAuMTAxNi9qLmJicmMuMjAxMy4xMS4wMTQ8L2VsZWN0cm9uaWMtcmVzb3Vy

Y2UtbnVtPjwvcmVjb3JkPjwvQ2l0ZT48Q2l0ZT48QXV0aG9yPlBlcmljbGVvdXM8L0F1dGhvcj48

WWVhcj4yMDA1PC9ZZWFyPjxSZWNOdW0+NDUwPC9SZWNOdW0+PHJlY29yZD48cmVjLW51bWJlcj40

NTA8L3JlYy1udW1iZXI+PGZvcmVpZ24ta2V5cz48a2V5IGFwcD0iRU4iIGRiLWlkPSI1c3owZTUy

enNzYXd4Y2VhMnhwcHc1NThmcno1enpzMmQwMnYiPjQ1MDwva2V5PjwvZm9yZWlnbi1rZXlzPjxy

ZWYtdHlwZSBuYW1lPSJKb3VybmFsIEFydGljbGUiPjE3PC9yZWYtdHlwZT48Y29udHJpYnV0b3Jz

PjxhdXRob3JzPjxhdXRob3I+UGVyaWNsZW91cywgTC4gTS48L2F1dGhvcj48YXV0aG9yPlJpY2hh

cmRzLCBKLjwvYXV0aG9yPjxhdXRob3I+RXBlbmV0b3MsIEEuIEEuPC9hdXRob3I+PGF1dGhvcj5D

b3VydGVuYXktTHVjaywgTi48L2F1dGhvcj48YXV0aG9yPkRlb25hcmFpbiwgTS4gUC48L2F1dGhv

cj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVzcz5EZXBhcnRtZW50IG9mIEJp

b2xvZ2ljYWwgU2NpZW5jZXMsIEltcGVyaWFsIENvbGxlZ2UgTG9uZG9uLCBVSy48L2F1dGgtYWRk

cmVzcz48dGl0bGVzPjx0aXRsZT5DaGFyYWN0ZXJpc2F0aW9uIGFuZCBpbnRlcm5hbGlzYXRpb24g

b2YgcmVjb21iaW5hbnQgaHVtYW5pc2VkIEhNRkctMSBhbnRpYm9kaWVzIGFnYWluc3QgTVVDMTwv

dGl0bGU+PHNlY29uZGFyeS10aXRsZT5CciBKIENhbmNlcjwvc2Vjb25kYXJ5LXRpdGxlPjxhbHQt

dGl0bGU+QnJpdGlzaCBqb3VybmFsIG9mIGNhbmNlcjwvYWx0LXRpdGxlPjwvdGl0bGVzPjxwZXJp

b2RpY2FsPjxmdWxsLXRpdGxlPkJyIEogQ2FuY2VyPC9mdWxsLXRpdGxlPjxhYmJyLTE+QnJpdGlz

aCBqb3VybmFsIG9mIGNhbmNlcjwvYWJici0xPjwvcGVyaW9kaWNhbD48YWx0LXBlcmlvZGljYWw+

PGZ1bGwtdGl0bGU+QnIgSiBDYW5jZXI8L2Z1bGwtdGl0bGU+PGFiYnItMT5Ccml0aXNoIGpvdXJu

YWwgb2YgY2FuY2VyPC9hYmJyLTE+PC9hbHQtcGVyaW9kaWNhbD48cGFnZXM+MTI1Ny02NjwvcGFn

ZXM+PHZvbHVtZT45Mzwvdm9sdW1lPjxudW1iZXI+MTE8L251bWJlcj48a2V5d29yZHM+PGtleXdv

cmQ+QW50aWJvZGllcywgTW9ub2Nsb25hbC8qaW1tdW5vbG9neTwva2V5d29yZD48a2V5d29yZD5C

aW5kaW5nIFNpdGVzLCBBbnRpYm9keTwva2V5d29yZD48a2V5d29yZD5CcmVhc3QgTmVvcGxhc21z

L3BhdGhvbG9neTwva2V5d29yZD48a2V5d29yZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdvcmQ+SW1t

dW5vZ2xvYnVsaW4gRmFiIEZyYWdtZW50cy9pbW11bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPktp

bmV0aWNzPC9rZXl3b3JkPjxrZXl3b3JkPk11Y2luLTEvKmltbXVub2xvZ3k8L2tleXdvcmQ+PGtl

eXdvcmQ+VHVtb3IgQ2VsbHMsIEN1bHR1cmVkPC9rZXl3b3JkPjwva2V5d29yZHM+PGRhdGVzPjx5

ZWFyPjIwMDU8L3llYXI+PHB1Yi1kYXRlcz48ZGF0ZT5Ob3YgMjg8L2RhdGU+PC9wdWItZGF0ZXM+

PC9kYXRlcz48aXNibj4wMDA3LTA5MjAgKFByaW50KSYjeEQ7MDAwNy0wOTIwIChMaW5raW5nKTwv

aXNibj48YWNjZXNzaW9uLW51bT4xNjI2NTM1MTwvYWNjZXNzaW9uLW51bT48dXJscz48cmVsYXRl

ZC11cmxzPjx1cmw+aHR0cDovL3d3dy5uY2JpLm5sbS5uaWguZ292L3B1Ym1lZC8xNjI2NTM1MTwv

dXJsPjwvcmVsYXRlZC11cmxzPjwvdXJscz48Y3VzdG9tMj4zMjE2MTExPC9jdXN0b20yPjxlbGVj

dHJvbmljLXJlc291cmNlLW51bT4xMC4xMDM4L3NqLmJqYy42NjAyODQ3PC9lbGVjdHJvbmljLXJl

c291cmNlLW51bT48L3JlY29yZD48L0NpdGU+PC9FbmROb3RlPgB=

ADDIN EN.CITE PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5MZWU8L0F1dGhvcj48WWVhcj4yMDEzPC9ZZWFyPjxSZWNO

dW0+NDQ5PC9SZWNOdW0+PERpc3BsYXlUZXh0PjxzdHlsZSBmYWNlPSJzdXBlcnNjcmlwdCI+MTAs

IDExPC9zdHlsZT48L0Rpc3BsYXlUZXh0PjxyZWNvcmQ+PHJlYy1udW1iZXI+NDQ5PC9yZWMtbnVt

YmVyPjxmb3JlaWduLWtleXM+PGtleSBhcHA9IkVOIiBkYi1pZD0iNXN6MGU1Mnpzc2F3eGNlYTJ4

cHB3NTU4ZnJ6NXp6czJkMDJ2Ij40NDk8L2tleT48L2ZvcmVpZ24ta2V5cz48cmVmLXR5cGUgbmFt

ZT0iSm91cm5hbCBBcnRpY2xlIj4xNzwvcmVmLXR5cGU+PGNvbnRyaWJ1dG9ycz48YXV0aG9ycz48

YXV0aG9yPkxlZSwgWS4gSC48L2F1dGhvcj48YXV0aG9yPklpamltYSwgTS48L2F1dGhvcj48YXV0

aG9yPkthZG8sIFkuPC9hdXRob3I+PGF1dGhvcj5NaXpvaGF0YSwgRS48L2F1dGhvcj48YXV0aG9y

Pklub3VlLCBULjwvYXV0aG9yPjxhdXRob3I+U3VnaXlhbWEsIEEuPC9hdXRob3I+PGF1dGhvcj5E

b2ksIEguPC9hdXRob3I+PGF1dGhvcj5TaGliYXNha2ksIFkuPC9hdXRob3I+PGF1dGhvcj5Lb2Rh

bWEsIFQuPC9hdXRob3I+PC9hdXRob3JzPjwvY29udHJpYnV0b3JzPjxhdXRoLWFkZHJlc3M+TGFi

b3JhdG9yeSBmb3IgU3lzdGVtcyBCaW9sb2d5IGFuZCBNZWRpY2luZSwgUmVzZWFyY2ggQ2VudGVy

IGZvciBBZHZhbmNlZCBTY2llbmNlIGFuZCBUZWNobm9sb2d5LCBUaGUgVW5pdmVyc2l0eSBvZiBU

b2t5bywgNC02LTEgS29tYWJhLCBNZWd1cm8ta3UsIFRva3lvIDE1My04OTA0LCBKYXBhbjsgRGVw

YXJ0bWVudCBvZiBMaWZlIFNjaWVuY2VzLCBHcmFkdWF0ZSBTY2hvb2wgb2YgQXJ0cyBhbmQgU2Np

ZW5jZXMsIFRoZSBVbml2ZXJzaXR5IG9mIFRva3lvLCAzLTgtMSBLb21hYmEsIE1lZ3Vyby1rdSwg

VG9reW8gMTUzLTg5MDIsIEphcGFuLjwvYXV0aC1hZGRyZXNzPjx0aXRsZXM+PHRpdGxlPkNvbnN0

cnVjdGlvbiBhbmQgY2hhcmFjdGVyaXphdGlvbiBvZiBmdW5jdGlvbmFsIGFudGktZXBpcmVndWxp

biBodW1hbml6ZWQgbW9ub2Nsb25hbCBhbnRpYm9kaWVzPC90aXRsZT48c2Vjb25kYXJ5LXRpdGxl

PkJpb2NoZW0gQmlvcGh5cyBSZXMgQ29tbXVuPC9zZWNvbmRhcnktdGl0bGU+PGFsdC10aXRsZT5C

aW9jaGVtaWNhbCBhbmQgYmlvcGh5c2ljYWwgcmVzZWFyY2ggY29tbXVuaWNhdGlvbnM8L2FsdC10

aXRsZT48L3RpdGxlcz48cGVyaW9kaWNhbD48ZnVsbC10aXRsZT5CaW9jaGVtIEJpb3BoeXMgUmVz

IENvbW11bjwvZnVsbC10aXRsZT48YWJici0xPkJpb2NoZW1pY2FsIGFuZCBiaW9waHlzaWNhbCBy

ZXNlYXJjaCBjb21tdW5pY2F0aW9uczwvYWJici0xPjwvcGVyaW9kaWNhbD48YWx0LXBlcmlvZGlj

YWw+PGZ1bGwtdGl0bGU+QmlvY2hlbSBCaW9waHlzIFJlcyBDb21tdW48L2Z1bGwtdGl0bGU+PGFi

YnItMT5CaW9jaGVtaWNhbCBhbmQgYmlvcGh5c2ljYWwgcmVzZWFyY2ggY29tbXVuaWNhdGlvbnM8

L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4xMDExLTc8L3BhZ2VzPjx2b2x1bWU+NDQx

PC92b2x1bWU+PG51bWJlcj40PC9udW1iZXI+PGtleXdvcmRzPjxrZXl3b3JkPkFtaW5vIEFjaWQg

U2VxdWVuY2U8L2tleXdvcmQ+PGtleXdvcmQ+QW50aWJvZGllcywgTW9ub2Nsb25hbCwgSHVtYW5p

emVkLypiaW9zeW50aGVzaXMvY2hlbWlzdHJ5L2ltbXVub2xvZ3k8L2tleXdvcmQ+PGtleXdvcmQ+

RXBpZGVybWFsIEdyb3d0aCBGYWN0b3IvYW50YWdvbmlzdHMgJmFtcDsgaW5oaWJpdG9ycy8qaW1t

dW5vbG9neTwva2V5d29yZD48a2V5d29yZD5IQ1QxMTYgQ2VsbHM8L2tleXdvcmQ+PGtleXdvcmQ+

SHVtYW5zPC9rZXl3b3JkPjxrZXl3b3JkPkltbXVub2dsb2J1bGluIFZhcmlhYmxlIFJlZ2lvbi9i

aW9zeW50aGVzaXMvY2hlbWlzdHJ5L2ltbXVub2xvZ3k8L2tleXdvcmQ+PGtleXdvcmQ+SnVya2F0

IENlbGxzPC9rZXl3b3JkPjxrZXl3b3JkPkx1Y2lmZXJhc2VzL2Jpb3N5bnRoZXNpczwva2V5d29y

ZD48a2V5d29yZD5Nb2RlbHMsIE1vbGVjdWxhcjwva2V5d29yZD48a2V5d29yZD5Nb2xlY3VsYXIg

U2VxdWVuY2UgRGF0YTwva2V5d29yZD48a2V5d29yZD5ORkFUQyBUcmFuc2NyaXB0aW9uIEZhY3Rv

cnMvbWV0YWJvbGlzbTwva2V5d29yZD48a2V5d29yZD5Qcm90ZWluIEVuZ2luZWVyaW5nPC9rZXl3

b3JkPjxrZXl3b3JkPlJlY2VwdG9ycywgSWdHL2Jpb3N5bnRoZXNpczwva2V5d29yZD48L2tleXdv

cmRzPjxkYXRlcz48eWVhcj4yMDEzPC95ZWFyPjxwdWItZGF0ZXM+PGRhdGU+Tm92IDI5PC9kYXRl

PjwvcHViLWRhdGVzPjwvZGF0ZXM+PGlzYm4+MTA5MC0yMTA0IChFbGVjdHJvbmljKSYjeEQ7MDAw

Ni0yOTFYIChMaW5raW5nKTwvaXNibj48YWNjZXNzaW9uLW51bT4yNDIzOTU0OTwvYWNjZXNzaW9u

LW51bT48dXJscz48cmVsYXRlZC11cmxzPjx1cmw+aHR0cDovL3d3dy5uY2JpLm5sbS5uaWguZ292

L3B1Ym1lZC8yNDIzOTU0OTwvdXJsPjwvcmVsYXRlZC11cmxzPjwvdXJscz48ZWxlY3Ryb25pYy1y

ZXNvdXJjZS1udW0+MTAuMTAxNi9qLmJicmMuMjAxMy4xMS4wMTQ8L2VsZWN0cm9uaWMtcmVzb3Vy

Y2UtbnVtPjwvcmVjb3JkPjwvQ2l0ZT48Q2l0ZT48QXV0aG9yPlBlcmljbGVvdXM8L0F1dGhvcj48

WWVhcj4yMDA1PC9ZZWFyPjxSZWNOdW0+NDUwPC9SZWNOdW0+PHJlY29yZD48cmVjLW51bWJlcj40

NTA8L3JlYy1udW1iZXI+PGZvcmVpZ24ta2V5cz48a2V5IGFwcD0iRU4iIGRiLWlkPSI1c3owZTUy

enNzYXd4Y2VhMnhwcHc1NThmcno1enpzMmQwMnYiPjQ1MDwva2V5PjwvZm9yZWlnbi1rZXlzPjxy

ZWYtdHlwZSBuYW1lPSJKb3VybmFsIEFydGljbGUiPjE3PC9yZWYtdHlwZT48Y29udHJpYnV0b3Jz

PjxhdXRob3JzPjxhdXRob3I+UGVyaWNsZW91cywgTC4gTS48L2F1dGhvcj48YXV0aG9yPlJpY2hh

cmRzLCBKLjwvYXV0aG9yPjxhdXRob3I+RXBlbmV0b3MsIEEuIEEuPC9hdXRob3I+PGF1dGhvcj5D

b3VydGVuYXktTHVjaywgTi48L2F1dGhvcj48YXV0aG9yPkRlb25hcmFpbiwgTS4gUC48L2F1dGhv

cj48L2F1dGhvcnM+PC9jb250cmlidXRvcnM+PGF1dGgtYWRkcmVzcz5EZXBhcnRtZW50IG9mIEJp

b2xvZ2ljYWwgU2NpZW5jZXMsIEltcGVyaWFsIENvbGxlZ2UgTG9uZG9uLCBVSy48L2F1dGgtYWRk

cmVzcz48dGl0bGVzPjx0aXRsZT5DaGFyYWN0ZXJpc2F0aW9uIGFuZCBpbnRlcm5hbGlzYXRpb24g

b2YgcmVjb21iaW5hbnQgaHVtYW5pc2VkIEhNRkctMSBhbnRpYm9kaWVzIGFnYWluc3QgTVVDMTwv

dGl0bGU+PHNlY29uZGFyeS10aXRsZT5CciBKIENhbmNlcjwvc2Vjb25kYXJ5LXRpdGxlPjxhbHQt

dGl0bGU+QnJpdGlzaCBqb3VybmFsIG9mIGNhbmNlcjwvYWx0LXRpdGxlPjwvdGl0bGVzPjxwZXJp

b2RpY2FsPjxmdWxsLXRpdGxlPkJyIEogQ2FuY2VyPC9mdWxsLXRpdGxlPjxhYmJyLTE+QnJpdGlz

aCBqb3VybmFsIG9mIGNhbmNlcjwvYWJici0xPjwvcGVyaW9kaWNhbD48YWx0LXBlcmlvZGljYWw+

PGZ1bGwtdGl0bGU+QnIgSiBDYW5jZXI8L2Z1bGwtdGl0bGU+PGFiYnItMT5Ccml0aXNoIGpvdXJu

YWwgb2YgY2FuY2VyPC9hYmJyLTE+PC9hbHQtcGVyaW9kaWNhbD48cGFnZXM+MTI1Ny02NjwvcGFn

ZXM+PHZvbHVtZT45Mzwvdm9sdW1lPjxudW1iZXI+MTE8L251bWJlcj48a2V5d29yZHM+PGtleXdv

cmQ+QW50aWJvZGllcywgTW9ub2Nsb25hbC8qaW1tdW5vbG9neTwva2V5d29yZD48a2V5d29yZD5C

aW5kaW5nIFNpdGVzLCBBbnRpYm9keTwva2V5d29yZD48a2V5d29yZD5CcmVhc3QgTmVvcGxhc21z

L3BhdGhvbG9neTwva2V5d29yZD48a2V5d29yZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdvcmQ+SW1t

dW5vZ2xvYnVsaW4gRmFiIEZyYWdtZW50cy9pbW11bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPktp

bmV0aWNzPC9rZXl3b3JkPjxrZXl3b3JkPk11Y2luLTEvKmltbXVub2xvZ3k8L2tleXdvcmQ+PGtl

eXdvcmQ+VHVtb3IgQ2VsbHMsIEN1bHR1cmVkPC9rZXl3b3JkPjwva2V5d29yZHM+PGRhdGVzPjx5

ZWFyPjIwMDU8L3llYXI+PHB1Yi1kYXRlcz48ZGF0ZT5Ob3YgMjg8L2RhdGU+PC9wdWItZGF0ZXM+

PC9kYXRlcz48aXNibj4wMDA3LTA5MjAgKFByaW50KSYjeEQ7MDAwNy0wOTIwIChMaW5raW5nKTwv

aXNibj48YWNjZXNzaW9uLW51bT4xNjI2NTM1MTwvYWNjZXNzaW9uLW51bT48dXJscz48cmVsYXRl

ZC11cmxzPjx1cmw+aHR0cDovL3d3dy5uY2JpLm5sbS5uaWguZ292L3B1Ym1lZC8xNjI2NTM1MTwv

dXJsPjwvcmVsYXRlZC11cmxzPjwvdXJscz48Y3VzdG9tMj4zMjE2MTExPC9jdXN0b20yPjxlbGVj

dHJvbmljLXJlc291cmNlLW51bT4xMC4xMDM4L3NqLmJqYy42NjAyODQ3PC9lbGVjdHJvbmljLXJl

c291cmNlLW51bT48L3JlY29yZD48L0NpdGU+PC9FbmROb3RlPgB=

ADDIN EN.CITE.DATA 10, 11 The apparent equilibrium dissociation constant (KD) including the avidity binding was also calculated using 1:1 Langmuir binding model.PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5QZXJpY2xlb3VzPC9BdXRob3I+PFllYXI+MjAwNTwvWWVh

cj48UmVjTnVtPjQ1MDwvUmVjTnVtPjxEaXNwbGF5VGV4dD48c3R5bGUgZmFjZT0ic3VwZXJzY3Jp

cHQiPjExLCAxMjwvc3R5bGU+PC9EaXNwbGF5VGV4dD48cmVjb3JkPjxyZWMtbnVtYmVyPjQ1MDwv

cmVjLW51bWJlcj48Zm9yZWlnbi1rZXlzPjxrZXkgYXBwPSJFTiIgZGItaWQ9IjVzejBlNTJ6c3Nh

d3hjZWEyeHBwdzU1OGZyejV6enMyZDAydiI+NDUwPC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10

eXBlIG5hbWU9IkpvdXJuYWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1

dGhvcnM+PGF1dGhvcj5QZXJpY2xlb3VzLCBMLiBNLjwvYXV0aG9yPjxhdXRob3I+UmljaGFyZHMs

IEouPC9hdXRob3I+PGF1dGhvcj5FcGVuZXRvcywgQS4gQS48L2F1dGhvcj48YXV0aG9yPkNvdXJ0

ZW5heS1MdWNrLCBOLjwvYXV0aG9yPjxhdXRob3I+RGVvbmFyYWluLCBNLiBQLjwvYXV0aG9yPjwv

YXV0aG9ycz48L2NvbnRyaWJ1dG9ycz48YXV0aC1hZGRyZXNzPkRlcGFydG1lbnQgb2YgQmlvbG9n

aWNhbCBTY2llbmNlcywgSW1wZXJpYWwgQ29sbGVnZSBMb25kb24sIFVLLjwvYXV0aC1hZGRyZXNz

Pjx0aXRsZXM+PHRpdGxlPkNoYXJhY3RlcmlzYXRpb24gYW5kIGludGVybmFsaXNhdGlvbiBvZiBy

ZWNvbWJpbmFudCBodW1hbmlzZWQgSE1GRy0xIGFudGlib2RpZXMgYWdhaW5zdCBNVUMxPC90aXRs

ZT48c2Vjb25kYXJ5LXRpdGxlPkJyIEogQ2FuY2VyPC9zZWNvbmRhcnktdGl0bGU+PGFsdC10aXRs

ZT5Ccml0aXNoIGpvdXJuYWwgb2YgY2FuY2VyPC9hbHQtdGl0bGU+PC90aXRsZXM+PHBlcmlvZGlj

YWw+PGZ1bGwtdGl0bGU+QnIgSiBDYW5jZXI8L2Z1bGwtdGl0bGU+PGFiYnItMT5Ccml0aXNoIGpv

dXJuYWwgb2YgY2FuY2VyPC9hYmJyLTE+PC9wZXJpb2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVs

bC10aXRsZT5CciBKIENhbmNlcjwvZnVsbC10aXRsZT48YWJici0xPkJyaXRpc2ggam91cm5hbCBv

ZiBjYW5jZXI8L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4xMjU3LTY2PC9wYWdlcz48

dm9sdW1lPjkzPC92b2x1bWU+PG51bWJlcj4xMTwvbnVtYmVyPjxrZXl3b3Jkcz48a2V5d29yZD5B

bnRpYm9kaWVzLCBNb25vY2xvbmFsLyppbW11bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPkJpbmRp

bmcgU2l0ZXMsIEFudGlib2R5PC9rZXl3b3JkPjxrZXl3b3JkPkJyZWFzdCBOZW9wbGFzbXMvcGF0

aG9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPkh1bWFuczwva2V5d29yZD48a2V5d29yZD5JbW11bm9n

bG9idWxpbiBGYWIgRnJhZ21lbnRzL2ltbXVub2xvZ3k8L2tleXdvcmQ+PGtleXdvcmQ+S2luZXRp

Y3M8L2tleXdvcmQ+PGtleXdvcmQ+TXVjaW4tMS8qaW1tdW5vbG9neTwva2V5d29yZD48a2V5d29y

ZD5UdW1vciBDZWxscywgQ3VsdHVyZWQ8L2tleXdvcmQ+PC9rZXl3b3Jkcz48ZGF0ZXM+PHllYXI+

MjAwNTwveWVhcj48cHViLWRhdGVzPjxkYXRlPk5vdiAyODwvZGF0ZT48L3B1Yi1kYXRlcz48L2Rh

dGVzPjxpc2JuPjAwMDctMDkyMCAoUHJpbnQpJiN4RDswMDA3LTA5MjAgKExpbmtpbmcpPC9pc2Ju

PjxhY2Nlc3Npb24tbnVtPjE2MjY1MzUxPC9hY2Nlc3Npb24tbnVtPjx1cmxzPjxyZWxhdGVkLXVy

bHM+PHVybD5odHRwOi8vd3d3Lm5jYmkubmxtLm5paC5nb3YvcHVibWVkLzE2MjY1MzUxPC91cmw+

PC9yZWxhdGVkLXVybHM+PC91cmxzPjxjdXN0b20yPjMyMTYxMTE8L2N1c3RvbTI+PGVsZWN0cm9u

aWMtcmVzb3VyY2UtbnVtPjEwLjEwMzgvc2ouYmpjLjY2MDI4NDc8L2VsZWN0cm9uaWMtcmVzb3Vy

Y2UtbnVtPjwvcmVjb3JkPjwvQ2l0ZT48Q2l0ZT48QXV0aG9yPlRyb2lzZTwvQXV0aG9yPjxZZWFy

PjIwMDg8L1llYXI+PFJlY051bT40NTE8L1JlY051bT48cmVjb3JkPjxyZWMtbnVtYmVyPjQ1MTwv

cmVjLW51bWJlcj48Zm9yZWlnbi1rZXlzPjxrZXkgYXBwPSJFTiIgZGItaWQ9IjVzejBlNTJ6c3Nh

d3hjZWEyeHBwdzU1OGZyejV6enMyZDAydiI+NDUxPC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10

eXBlIG5hbWU9IkpvdXJuYWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1

dGhvcnM+PGF1dGhvcj5Ucm9pc2UsIEYuPC9hdXRob3I+PGF1dGhvcj5DYWZhcm8sIFYuPC9hdXRo

b3I+PGF1dGhvcj5HaWFuY29sYSwgQy48L2F1dGhvcj48YXV0aG9yPkQmYXBvcztBbGVzc2lvLCBH

LjwvYXV0aG9yPjxhdXRob3I+RGUgTG9yZW56bywgQy48L2F1dGhvcj48L2F1dGhvcnM+PC9jb250

cmlidXRvcnM+PGF1dGgtYWRkcmVzcz5EaXBhcnRpbWVudG8gZGkgQmlvbG9naWEgU3RydXR0dXJh

bGUgZSBGdW56aW9uYWxlLCBVbml2ZXJzaXRhIGRpIE5hcG9saSBGZWRlcmljbyBJSSwgSXRhbHku

PC9hdXRoLWFkZHJlc3M+PHRpdGxlcz48dGl0bGU+RGlmZmVyZW50aWFsIGJpbmRpbmcgb2YgaHVt

YW4gaW1tdW5vYWdlbnRzIGFuZCBIZXJjZXB0aW4gdG8gdGhlIEVyYkIyIHJlY2VwdG9yPC90aXRs

ZT48c2Vjb25kYXJ5LXRpdGxlPkZFQlMgSjwvc2Vjb25kYXJ5LXRpdGxlPjxhbHQtdGl0bGU+VGhl

IEZFQlMgam91cm5hbDwvYWx0LXRpdGxlPjwvdGl0bGVzPjxwZXJpb2RpY2FsPjxmdWxsLXRpdGxl

PkZFQlMgSjwvZnVsbC10aXRsZT48YWJici0xPlRoZSBGRUJTIGpvdXJuYWw8L2FiYnItMT48L3Bl

cmlvZGljYWw+PGFsdC1wZXJpb2RpY2FsPjxmdWxsLXRpdGxlPkZFQlMgSjwvZnVsbC10aXRsZT48

YWJici0xPlRoZSBGRUJTIGpvdXJuYWw8L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz40

OTY3LTc5PC9wYWdlcz48dm9sdW1lPjI3NTwvdm9sdW1lPjxudW1iZXI+MjA8L251bWJlcj48a2V5

d29yZHM+PGtleXdvcmQ+QW50aWJvZGllcy9nZW5ldGljcy9waGFybWFjb2xvZ3k8L2tleXdvcmQ+

PGtleXdvcmQ+QW50aWJvZGllcywgTW9ub2Nsb25hbC9waGFybWFjb2xvZ3k8L2tleXdvcmQ+PGtl

eXdvcmQ+QW50aWJvZGllcywgTW9ub2Nsb25hbCwgSHVtYW5pemVkPC9rZXl3b3JkPjxrZXl3b3Jk

PkFudGluZW9wbGFzdGljIEFnZW50cy8qcGhhcm1hY29sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPkJy

ZWFzdCBOZW9wbGFzbXMvZHJ1ZyB0aGVyYXB5L3BhdGhvbG9neTwva2V5d29yZD48a2V5d29yZD5G

ZW1hbGU8L2tleXdvcmQ+PGtleXdvcmQ+SHVtYW5zPC9rZXl3b3JkPjxrZXl3b3JkPkltbXVub2ds

b2J1bGluIEZyYWdtZW50cy9nZW5ldGljcy9waGFybWFjb2xvZ3k8L2tleXdvcmQ+PGtleXdvcmQ+

SW1tdW5vdGhlcmFweS8qbWV0aG9kczwva2V5d29yZD48a2V5d29yZD5Qcm90ZWluIEJpbmRpbmc8

L2tleXdvcmQ+PGtleXdvcmQ+UHJvdGVpbiBFbmdpbmVlcmluZy8qbWV0aG9kczwva2V5d29yZD48

a2V5d29yZD5SZWNlcHRvciwgZXJiQi0yLyppbW11bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPlJl

Y29tYmluYW50IEZ1c2lvbiBQcm90ZWlucy8qdGhlcmFwZXV0aWMgdXNlPC9rZXl3b3JkPjxrZXl3

b3JkPlR1bW9yIENlbGxzLCBDdWx0dXJlZDwva2V5d29yZD48L2tleXdvcmRzPjxkYXRlcz48eWVh

cj4yMDA4PC95ZWFyPjxwdWItZGF0ZXM+PGRhdGU+T2N0PC9kYXRlPjwvcHViLWRhdGVzPjwvZGF0

ZXM+PGlzYm4+MTc0Mi00NjU4IChFbGVjdHJvbmljKSYjeEQ7MTc0Mi00NjRYIChMaW5raW5nKTwv

aXNibj48YWNjZXNzaW9uLW51bT4xODc5NTk1MDwvYWNjZXNzaW9uLW51bT48dXJscz48cmVsYXRl

ZC11cmxzPjx1cmw+aHR0cDovL3d3dy5uY2JpLm5sbS5uaWguZ292L3B1Ym1lZC8xODc5NTk1MDwv

dXJsPjwvcmVsYXRlZC11cmxzPjwvdXJscz48ZWxlY3Ryb25pYy1yZXNvdXJjZS1udW0+MTAuMTEx

MS9qLjE3NDItNDY1OC4yMDA4LjA2NjI1Lng8L2VsZWN0cm9uaWMtcmVzb3VyY2UtbnVtPjwvcmVj

b3JkPjwvQ2l0ZT48L0VuZE5vdGU+AG==

ADDIN EN.CITE PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5QZXJpY2xlb3VzPC9BdXRob3I+PFllYXI+MjAwNTwvWWVh

cj48UmVjTnVtPjQ1MDwvUmVjTnVtPjxEaXNwbGF5VGV4dD48c3R5bGUgZmFjZT0ic3VwZXJzY3Jp

cHQiPjExLCAxMjwvc3R5bGU+PC9EaXNwbGF5VGV4dD48cmVjb3JkPjxyZWMtbnVtYmVyPjQ1MDwv

cmVjLW51bWJlcj48Zm9yZWlnbi1rZXlzPjxrZXkgYXBwPSJFTiIgZGItaWQ9IjVzejBlNTJ6c3Nh

d3hjZWEyeHBwdzU1OGZyejV6enMyZDAydiI+NDUwPC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10

eXBlIG5hbWU9IkpvdXJuYWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1

dGhvcnM+PGF1dGhvcj5QZXJpY2xlb3VzLCBMLiBNLjwvYXV0aG9yPjxhdXRob3I+UmljaGFyZHMs

IEouPC9hdXRob3I+PGF1dGhvcj5FcGVuZXRvcywgQS4gQS48L2F1dGhvcj48YXV0aG9yPkNvdXJ0

ZW5heS1MdWNrLCBOLjwvYXV0aG9yPjxhdXRob3I+RGVvbmFyYWluLCBNLiBQLjwvYXV0aG9yPjwv

YXV0aG9ycz48L2NvbnRyaWJ1dG9ycz48YXV0aC1hZGRyZXNzPkRlcGFydG1lbnQgb2YgQmlvbG9n

aWNhbCBTY2llbmNlcywgSW1wZXJpYWwgQ29sbGVnZSBMb25kb24sIFVLLjwvYXV0aC1hZGRyZXNz

Pjx0aXRsZXM+PHRpdGxlPkNoYXJhY3RlcmlzYXRpb24gYW5kIGludGVybmFsaXNhdGlvbiBvZiBy

ZWNvbWJpbmFudCBodW1hbmlzZWQgSE1GRy0xIGFudGlib2RpZXMgYWdhaW5zdCBNVUMxPC90aXRs

ZT48c2Vjb25kYXJ5LXRpdGxlPkJyIEogQ2FuY2VyPC9zZWNvbmRhcnktdGl0bGU+PGFsdC10aXRs

ZT5Ccml0aXNoIGpvdXJuYWwgb2YgY2FuY2VyPC9hbHQtdGl0bGU+PC90aXRsZXM+PHBlcmlvZGlj

YWw+PGZ1bGwtdGl0bGU+QnIgSiBDYW5jZXI8L2Z1bGwtdGl0bGU+PGFiYnItMT5Ccml0aXNoIGpv

dXJuYWwgb2YgY2FuY2VyPC9hYmJyLTE+PC9wZXJpb2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVs

bC10aXRsZT5CciBKIENhbmNlcjwvZnVsbC10aXRsZT48YWJici0xPkJyaXRpc2ggam91cm5hbCBv

ZiBjYW5jZXI8L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4xMjU3LTY2PC9wYWdlcz48

dm9sdW1lPjkzPC92b2x1bWU+PG51bWJlcj4xMTwvbnVtYmVyPjxrZXl3b3Jkcz48a2V5d29yZD5B

bnRpYm9kaWVzLCBNb25vY2xvbmFsLyppbW11bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPkJpbmRp

bmcgU2l0ZXMsIEFudGlib2R5PC9rZXl3b3JkPjxrZXl3b3JkPkJyZWFzdCBOZW9wbGFzbXMvcGF0

aG9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPkh1bWFuczwva2V5d29yZD48a2V5d29yZD5JbW11bm9n

bG9idWxpbiBGYWIgRnJhZ21lbnRzL2ltbXVub2xvZ3k8L2tleXdvcmQ+PGtleXdvcmQ+S2luZXRp

Y3M8L2tleXdvcmQ+PGtleXdvcmQ+TXVjaW4tMS8qaW1tdW5vbG9neTwva2V5d29yZD48a2V5d29y

ZD5UdW1vciBDZWxscywgQ3VsdHVyZWQ8L2tleXdvcmQ+PC9rZXl3b3Jkcz48ZGF0ZXM+PHllYXI+

MjAwNTwveWVhcj48cHViLWRhdGVzPjxkYXRlPk5vdiAyODwvZGF0ZT48L3B1Yi1kYXRlcz48L2Rh

dGVzPjxpc2JuPjAwMDctMDkyMCAoUHJpbnQpJiN4RDswMDA3LTA5MjAgKExpbmtpbmcpPC9pc2Ju

PjxhY2Nlc3Npb24tbnVtPjE2MjY1MzUxPC9hY2Nlc3Npb24tbnVtPjx1cmxzPjxyZWxhdGVkLXVy

bHM+PHVybD5odHRwOi8vd3d3Lm5jYmkubmxtLm5paC5nb3YvcHVibWVkLzE2MjY1MzUxPC91cmw+

PC9yZWxhdGVkLXVybHM+PC91cmxzPjxjdXN0b20yPjMyMTYxMTE8L2N1c3RvbTI+PGVsZWN0cm9u

aWMtcmVzb3VyY2UtbnVtPjEwLjEwMzgvc2ouYmpjLjY2MDI4NDc8L2VsZWN0cm9uaWMtcmVzb3Vy

Y2UtbnVtPjwvcmVjb3JkPjwvQ2l0ZT48Q2l0ZT48QXV0aG9yPlRyb2lzZTwvQXV0aG9yPjxZZWFy

PjIwMDg8L1llYXI+PFJlY051bT40NTE8L1JlY051bT48cmVjb3JkPjxyZWMtbnVtYmVyPjQ1MTwv

cmVjLW51bWJlcj48Zm9yZWlnbi1rZXlzPjxrZXkgYXBwPSJFTiIgZGItaWQ9IjVzejBlNTJ6c3Nh

d3hjZWEyeHBwdzU1OGZyejV6enMyZDAydiI+NDUxPC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10

eXBlIG5hbWU9IkpvdXJuYWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1

dGhvcnM+PGF1dGhvcj5Ucm9pc2UsIEYuPC9hdXRob3I+PGF1dGhvcj5DYWZhcm8sIFYuPC9hdXRo

b3I+PGF1dGhvcj5HaWFuY29sYSwgQy48L2F1dGhvcj48YXV0aG9yPkQmYXBvcztBbGVzc2lvLCBH

LjwvYXV0aG9yPjxhdXRob3I+RGUgTG9yZW56bywgQy48L2F1dGhvcj48L2F1dGhvcnM+PC9jb250

cmlidXRvcnM+PGF1dGgtYWRkcmVzcz5EaXBhcnRpbWVudG8gZGkgQmlvbG9naWEgU3RydXR0dXJh

bGUgZSBGdW56aW9uYWxlLCBVbml2ZXJzaXRhIGRpIE5hcG9saSBGZWRlcmljbyBJSSwgSXRhbHku

PC9hdXRoLWFkZHJlc3M+PHRpdGxlcz48dGl0bGU+RGlmZmVyZW50aWFsIGJpbmRpbmcgb2YgaHVt

YW4gaW1tdW5vYWdlbnRzIGFuZCBIZXJjZXB0aW4gdG8gdGhlIEVyYkIyIHJlY2VwdG9yPC90aXRs

ZT48c2Vjb25kYXJ5LXRpdGxlPkZFQlMgSjwvc2Vjb25kYXJ5LXRpdGxlPjxhbHQtdGl0bGU+VGhl

IEZFQlMgam91cm5hbDwvYWx0LXRpdGxlPjwvdGl0bGVzPjxwZXJpb2RpY2FsPjxmdWxsLXRpdGxl

PkZFQlMgSjwvZnVsbC10aXRsZT48YWJici0xPlRoZSBGRUJTIGpvdXJuYWw8L2FiYnItMT48L3Bl

cmlvZGljYWw+PGFsdC1wZXJpb2RpY2FsPjxmdWxsLXRpdGxlPkZFQlMgSjwvZnVsbC10aXRsZT48

YWJici0xPlRoZSBGRUJTIGpvdXJuYWw8L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz40

OTY3LTc5PC9wYWdlcz48dm9sdW1lPjI3NTwvdm9sdW1lPjxudW1iZXI+MjA8L251bWJlcj48a2V5

d29yZHM+PGtleXdvcmQ+QW50aWJvZGllcy9nZW5ldGljcy9waGFybWFjb2xvZ3k8L2tleXdvcmQ+

PGtleXdvcmQ+QW50aWJvZGllcywgTW9ub2Nsb25hbC9waGFybWFjb2xvZ3k8L2tleXdvcmQ+PGtl

eXdvcmQ+QW50aWJvZGllcywgTW9ub2Nsb25hbCwgSHVtYW5pemVkPC9rZXl3b3JkPjxrZXl3b3Jk

PkFudGluZW9wbGFzdGljIEFnZW50cy8qcGhhcm1hY29sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPkJy

ZWFzdCBOZW9wbGFzbXMvZHJ1ZyB0aGVyYXB5L3BhdGhvbG9neTwva2V5d29yZD48a2V5d29yZD5G

ZW1hbGU8L2tleXdvcmQ+PGtleXdvcmQ+SHVtYW5zPC9rZXl3b3JkPjxrZXl3b3JkPkltbXVub2ds

b2J1bGluIEZyYWdtZW50cy9nZW5ldGljcy9waGFybWFjb2xvZ3k8L2tleXdvcmQ+PGtleXdvcmQ+

SW1tdW5vdGhlcmFweS8qbWV0aG9kczwva2V5d29yZD48a2V5d29yZD5Qcm90ZWluIEJpbmRpbmc8

L2tleXdvcmQ+PGtleXdvcmQ+UHJvdGVpbiBFbmdpbmVlcmluZy8qbWV0aG9kczwva2V5d29yZD48

a2V5d29yZD5SZWNlcHRvciwgZXJiQi0yLyppbW11bm9sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPlJl

Y29tYmluYW50IEZ1c2lvbiBQcm90ZWlucy8qdGhlcmFwZXV0aWMgdXNlPC9rZXl3b3JkPjxrZXl3

b3JkPlR1bW9yIENlbGxzLCBDdWx0dXJlZDwva2V5d29yZD48L2tleXdvcmRzPjxkYXRlcz48eWVh

cj4yMDA4PC95ZWFyPjxwdWItZGF0ZXM+PGRhdGU+T2N0PC9kYXRlPjwvcHViLWRhdGVzPjwvZGF0

ZXM+PGlzYm4+MTc0Mi00NjU4IChFbGVjdHJvbmljKSYjeEQ7MTc0Mi00NjRYIChMaW5raW5nKTwv

aXNibj48YWNjZXNzaW9uLW51bT4xODc5NTk1MDwvYWNjZXNzaW9uLW51bT48dXJscz48cmVsYXRl

ZC11cmxzPjx1cmw+aHR0cDovL3d3dy5uY2JpLm5sbS5uaWguZ292L3B1Ym1lZC8xODc5NTk1MDwv

dXJsPjwvcmVsYXRlZC11cmxzPjwvdXJscz48ZWxlY3Ryb25pYy1yZXNvdXJjZS1udW0+MTAuMTEx

MS9qLjE3NDItNDY1OC4yMDA4LjA2NjI1Lng8L2VsZWN0cm9uaWMtcmVzb3VyY2UtbnVtPjwvcmVj

b3JkPjwvQ2l0ZT48L0VuZE5vdGU+AG==

ADDIN EN.CITE.DATA 11, 12DNA hydrolyzing assayAs a substrate, supercoiled plasmid pUC19 were purified using a plasmid miniprep kit (Nanohelix, FPL200). DNA hydrolyzing experiments were initiated by mixing proteins such as m3D8 scFv (0.1 ~ 0.5 ?M) and indicated cytotransmabs (0.1 ?M) with DNA substrates (2.2 nM) in a TBS buffer (50 mM Tris-Cl, pH 7.4, 50 mM NaCl) containing either 2 mM MgCl2 or 50 mM EDTA, as previously described previously.PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5LaW08L0F1dGhvcj48WWVhcj4yMDA5PC9ZZWFyPjxSZWNO

dW0+NDU8L1JlY051bT48RGlzcGxheVRleHQ+PHN0eWxlIGZhY2U9InN1cGVyc2NyaXB0Ij4zLCA2

PC9zdHlsZT48L0Rpc3BsYXlUZXh0PjxyZWNvcmQ+PHJlYy1udW1iZXI+NDU8L3JlYy1udW1iZXI+

PGZvcmVpZ24ta2V5cz48a2V5IGFwcD0iRU4iIGRiLWlkPSI1c3owZTUyenNzYXd4Y2VhMnhwcHc1

NThmcno1enpzMmQwMnYiPjQ1PC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10eXBlIG5hbWU9Ikpv

dXJuYWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1dGhvcnM+PGF1dGhv

cj5LaW0sIEQuIFMuPC9hdXRob3I+PGF1dGhvcj5MZWUsIFMuIEguPC9hdXRob3I+PGF1dGhvcj5L

aW0sIEouIFMuPC9hdXRob3I+PGF1dGhvcj5MZWUsIFMuIEMuPC9hdXRob3I+PGF1dGhvcj5Ld29u

LCBNLiBILjwvYXV0aG9yPjxhdXRob3I+S2ltLCBZLiBTLjwvYXV0aG9yPjwvYXV0aG9ycz48L2Nv

bnRyaWJ1dG9ycz48YXV0aC1hZGRyZXNzPkRlcGFydG1lbnQgb2YgTW9sZWN1bGFyIFNjaWVuY2Ug

YW5kIFRlY2hub2xvZ3ksIEFqb3UgVW5pdmVyc2l0eSwgU2FuIDUsIFdvbmNoZW9uLWRvbmcsIFll

b25ndG9uZy1ndSwgU3V3b24gNDQzLTc0OSwgUmVwdWJsaWMgb2YgS29yZWEuPC9hdXRoLWFkZHJl

c3M+PHRpdGxlcz48dGl0bGU+R2VuZXJhdGlvbiBvZiBodW1hbml6ZWQgYW50aS1ETkEgaHlkcm9s

eXppbmcgY2F0YWx5dGljIGFudGlib2RpZXMgYnkgY29tcGxlbWVudGFyaXR5IGRldGVybWluaW5n

IHJlZ2lvbiBncmFmdGluZzwvdGl0bGU+PHNlY29uZGFyeS10aXRsZT5CaW9jaGVtIEJpb3BoeXMg

UmVzIENvbW11bjwvc2Vjb25kYXJ5LXRpdGxlPjwvdGl0bGVzPjxwZXJpb2RpY2FsPjxmdWxsLXRp

dGxlPkJpb2NoZW0gQmlvcGh5cyBSZXMgQ29tbXVuPC9mdWxsLXRpdGxlPjxhYmJyLTE+QmlvY2hl

bWljYWwgYW5kIGJpb3BoeXNpY2FsIHJlc2VhcmNoIGNvbW11bmljYXRpb25zPC9hYmJyLTE+PC9w

ZXJpb2RpY2FsPjxwYWdlcz4zMTQtODwvcGFnZXM+PHZvbHVtZT4zNzk8L3ZvbHVtZT48bnVtYmVy

PjI8L251bWJlcj48ZGF0ZXM+PHllYXI+MjAwOTwveWVhcj48cHViLWRhdGVzPjxkYXRlPkZlYiA2

PC9kYXRlPjwvcHViLWRhdGVzPjwvZGF0ZXM+PGFjY2Vzc2lvbi1udW0+MTkxMDMxNzE8L2FjY2Vz

c2lvbi1udW0+PHVybHM+PHJlbGF0ZWQtdXJscz48dXJsPmh0dHA6Ly93d3cubmNiaS5ubG0ubmlo

Lmdvdi9lbnRyZXovcXVlcnkuZmNnaT9jbWQ9UmV0cmlldmUmYW1wO2RiPVB1Yk1lZCZhbXA7ZG9w

dD1DaXRhdGlvbiZhbXA7bGlzdF91aWRzPTE5MTAzMTcxIDwvdXJsPjwvcmVsYXRlZC11cmxzPjwv

dXJscz48L3JlY29yZD48L0NpdGU+PENpdGU+PEF1dGhvcj5MZWU8L0F1dGhvcj48WWVhcj4yMDEw

PC9ZZWFyPjxSZWNOdW0+MjI8L1JlY051bT48cmVjb3JkPjxyZWMtbnVtYmVyPjIyPC9yZWMtbnVt

YmVyPjxmb3JlaWduLWtleXM+PGtleSBhcHA9IkVOIiBkYi1pZD0iNXN6MGU1Mnpzc2F3eGNlYTJ4

cHB3NTU4ZnJ6NXp6czJkMDJ2Ij4yMjwva2V5PjwvZm9yZWlnbi1rZXlzPjxyZWYtdHlwZSBuYW1l

PSJKb3VybmFsIEFydGljbGUiPjE3PC9yZWYtdHlwZT48Y29udHJpYnV0b3JzPjxhdXRob3JzPjxh

dXRob3I+TGVlLCBXLiBSLjwvYXV0aG9yPjxhdXRob3I+SmFuZywgSi4gWS48L2F1dGhvcj48YXV0

aG9yPktpbSwgSi4gUy48L2F1dGhvcj48YXV0aG9yPkt3b24sIE0uIEguPC9hdXRob3I+PGF1dGhv

cj5LaW0sIFkuIFMuPC9hdXRob3I+PC9hdXRob3JzPjwvY29udHJpYnV0b3JzPjxhdXRoLWFkZHJl

c3M+RGVwYXJ0bWVudCBvZiBNb2xlY3VsYXIgU2NpZW5jZSBhbmQgVGVjaG5vbG9neSwgQWpvdSBV

bml2ZXJzaXR5LCBTdXdvbiA0NDMtNzQ5LCBLb3JlYS48L2F1dGgtYWRkcmVzcz48dGl0bGVzPjx0

aXRsZT5HZW5lIHNpbGVuY2luZyBieSBjZWxsLXBlbmV0cmF0aW5nLCBzZXF1ZW5jZS1zZWxlY3Rp

dmUgYW5kIG51Y2xlaWMtYWNpZCBoeWRyb2x5emluZyBhbnRpYm9kaWVzPC90aXRsZT48c2Vjb25k

YXJ5LXRpdGxlPk51Y2xlaWMgQWNpZHMgUmVzPC9zZWNvbmRhcnktdGl0bGU+PGFsdC10aXRsZT5O

dWNsZWljIGFjaWRzIHJlc2VhcmNoPC9hbHQtdGl0bGU+PC90aXRsZXM+PHBlcmlvZGljYWw+PGZ1

bGwtdGl0bGU+TnVjbGVpYyBBY2lkcyBSZXM8L2Z1bGwtdGl0bGU+PGFiYnItMT5OdWNsZWljIGFj

aWRzIHJlc2VhcmNoPC9hYmJyLTE+PC9wZXJpb2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVsbC10

aXRsZT5OdWNsZWljIEFjaWRzIFJlczwvZnVsbC10aXRsZT48YWJici0xPk51Y2xlaWMgYWNpZHMg

cmVzZWFyY2g8L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4xNTk2LTYwOTwvcGFnZXM+

PHZvbHVtZT4zODwvdm9sdW1lPjxudW1iZXI+NTwvbnVtYmVyPjxlZGl0aW9uPjIwMDkvMTIvMTc8

L2VkaXRpb24+PGtleXdvcmRzPjxrZXl3b3JkPkFudGlib2RpZXMsIENhdGFseXRpYy9jaGVtaXN0

cnkvKm1ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+QXBvcHRvc2lzPC9rZXl3b3JkPjxrZXl3

b3JkPkJhc2UgU2VxdWVuY2U8L2tleXdvcmQ+PGtleXdvcmQ+Q2VsbCBMaW5lLCBUdW1vcjwva2V5

d29yZD48a2V5d29yZD5DeXRvc29sL21ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+RE5BLCBT

aW5nbGUtU3RyYW5kZWQvbWV0YWJvbGlzbTwva2V5d29yZD48a2V5d29yZD5HZW5lIEtub2NrZG93

biBUZWNobmlxdWVzPC9rZXl3b3JkPjxrZXl3b3JkPkdlbmUgTGlicmFyeTwva2V5d29yZD48a2V5

d29yZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdvcmQ+SHlkcm9seXNpczwva2V5d29yZD48a2V5d29y

ZD5JbW11bm9nbG9idWxpbiBMaWdodCBDaGFpbnMvY2hlbWlzdHJ5PC9rZXl3b3JkPjxrZXl3b3Jk

PkltbXVub2dsb2J1bGluIFZhcmlhYmxlIFJlZ2lvbi9jaGVtaXN0cnk8L2tleXdvcmQ+PGtleXdv

cmQ+KlJOQSBJbnRlcmZlcmVuY2U8L2tleXdvcmQ+PGtleXdvcmQ+Uk5BLCBNZXNzZW5nZXIvKm1l

dGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+UmVjZXB0b3IsIGVyYkItMi9hbnRhZ29uaXN0cyAm

YW1wOyBpbmhpYml0b3JzL2dlbmV0aWNzL21ldGFib2xpc208L2tleXdvcmQ+PC9rZXl3b3Jkcz48

ZGF0ZXM+PHllYXI+MjAxMDwveWVhcj48cHViLWRhdGVzPjxkYXRlPk1hcjwvZGF0ZT48L3B1Yi1k

YXRlcz48L2RhdGVzPjxpc2JuPjEzNjItNDk2MiAoRWxlY3Ryb25pYykmI3hEOzAzMDUtMTA0OCAo

TGlua2luZyk8L2lzYm4+PGFjY2Vzc2lvbi1udW0+MjAwMDc2MDI8L2FjY2Vzc2lvbi1udW0+PHdv

cmstdHlwZT5SZXNlYXJjaCBTdXBwb3J0LCBOb24tVS5TLiBHb3YmYXBvczt0PC93b3JrLXR5cGU+

PHVybHM+PHJlbGF0ZWQtdXJscz48dXJsPmh0dHA6Ly93d3cubmNiaS5ubG0ubmloLmdvdi9wdWJt

ZWQvMjAwMDc2MDI8L3VybD48L3JlbGF0ZWQtdXJscz48L3VybHM+PGN1c3RvbTI+MjgzNjU3Mjwv

Y3VzdG9tMj48ZWxlY3Ryb25pYy1yZXNvdXJjZS1udW0+MTAuMTA5My9uYXIvZ2twMTE0NTwvZWxl

Y3Ryb25pYy1yZXNvdXJjZS1udW0+PGxhbmd1YWdlPmVuZzwvbGFuZ3VhZ2U+PC9yZWNvcmQ+PC9D

aXRlPjwvRW5kTm90ZT4A

ADDIN EN.CITE PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5LaW08L0F1dGhvcj48WWVhcj4yMDA5PC9ZZWFyPjxSZWNO

dW0+NDU8L1JlY051bT48RGlzcGxheVRleHQ+PHN0eWxlIGZhY2U9InN1cGVyc2NyaXB0Ij4zLCA2

PC9zdHlsZT48L0Rpc3BsYXlUZXh0PjxyZWNvcmQ+PHJlYy1udW1iZXI+NDU8L3JlYy1udW1iZXI+

PGZvcmVpZ24ta2V5cz48a2V5IGFwcD0iRU4iIGRiLWlkPSI1c3owZTUyenNzYXd4Y2VhMnhwcHc1

NThmcno1enpzMmQwMnYiPjQ1PC9rZXk+PC9mb3JlaWduLWtleXM+PHJlZi10eXBlIG5hbWU9Ikpv

dXJuYWwgQXJ0aWNsZSI+MTc8L3JlZi10eXBlPjxjb250cmlidXRvcnM+PGF1dGhvcnM+PGF1dGhv

cj5LaW0sIEQuIFMuPC9hdXRob3I+PGF1dGhvcj5MZWUsIFMuIEguPC9hdXRob3I+PGF1dGhvcj5L

aW0sIEouIFMuPC9hdXRob3I+PGF1dGhvcj5MZWUsIFMuIEMuPC9hdXRob3I+PGF1dGhvcj5Ld29u

LCBNLiBILjwvYXV0aG9yPjxhdXRob3I+S2ltLCBZLiBTLjwvYXV0aG9yPjwvYXV0aG9ycz48L2Nv

bnRyaWJ1dG9ycz48YXV0aC1hZGRyZXNzPkRlcGFydG1lbnQgb2YgTW9sZWN1bGFyIFNjaWVuY2Ug

YW5kIFRlY2hub2xvZ3ksIEFqb3UgVW5pdmVyc2l0eSwgU2FuIDUsIFdvbmNoZW9uLWRvbmcsIFll

b25ndG9uZy1ndSwgU3V3b24gNDQzLTc0OSwgUmVwdWJsaWMgb2YgS29yZWEuPC9hdXRoLWFkZHJl

c3M+PHRpdGxlcz48dGl0bGU+R2VuZXJhdGlvbiBvZiBodW1hbml6ZWQgYW50aS1ETkEgaHlkcm9s

eXppbmcgY2F0YWx5dGljIGFudGlib2RpZXMgYnkgY29tcGxlbWVudGFyaXR5IGRldGVybWluaW5n

IHJlZ2lvbiBncmFmdGluZzwvdGl0bGU+PHNlY29uZGFyeS10aXRsZT5CaW9jaGVtIEJpb3BoeXMg

UmVzIENvbW11bjwvc2Vjb25kYXJ5LXRpdGxlPjwvdGl0bGVzPjxwZXJpb2RpY2FsPjxmdWxsLXRp

dGxlPkJpb2NoZW0gQmlvcGh5cyBSZXMgQ29tbXVuPC9mdWxsLXRpdGxlPjxhYmJyLTE+QmlvY2hl

bWljYWwgYW5kIGJpb3BoeXNpY2FsIHJlc2VhcmNoIGNvbW11bmljYXRpb25zPC9hYmJyLTE+PC9w

ZXJpb2RpY2FsPjxwYWdlcz4zMTQtODwvcGFnZXM+PHZvbHVtZT4zNzk8L3ZvbHVtZT48bnVtYmVy

PjI8L251bWJlcj48ZGF0ZXM+PHllYXI+MjAwOTwveWVhcj48cHViLWRhdGVzPjxkYXRlPkZlYiA2

PC9kYXRlPjwvcHViLWRhdGVzPjwvZGF0ZXM+PGFjY2Vzc2lvbi1udW0+MTkxMDMxNzE8L2FjY2Vz

c2lvbi1udW0+PHVybHM+PHJlbGF0ZWQtdXJscz48dXJsPmh0dHA6Ly93d3cubmNiaS5ubG0ubmlo

Lmdvdi9lbnRyZXovcXVlcnkuZmNnaT9jbWQ9UmV0cmlldmUmYW1wO2RiPVB1Yk1lZCZhbXA7ZG9w

dD1DaXRhdGlvbiZhbXA7bGlzdF91aWRzPTE5MTAzMTcxIDwvdXJsPjwvcmVsYXRlZC11cmxzPjwv

dXJscz48L3JlY29yZD48L0NpdGU+PENpdGU+PEF1dGhvcj5MZWU8L0F1dGhvcj48WWVhcj4yMDEw

PC9ZZWFyPjxSZWNOdW0+MjI8L1JlY051bT48cmVjb3JkPjxyZWMtbnVtYmVyPjIyPC9yZWMtbnVt

YmVyPjxmb3JlaWduLWtleXM+PGtleSBhcHA9IkVOIiBkYi1pZD0iNXN6MGU1Mnpzc2F3eGNlYTJ4

cHB3NTU4ZnJ6NXp6czJkMDJ2Ij4yMjwva2V5PjwvZm9yZWlnbi1rZXlzPjxyZWYtdHlwZSBuYW1l

PSJKb3VybmFsIEFydGljbGUiPjE3PC9yZWYtdHlwZT48Y29udHJpYnV0b3JzPjxhdXRob3JzPjxh

dXRob3I+TGVlLCBXLiBSLjwvYXV0aG9yPjxhdXRob3I+SmFuZywgSi4gWS48L2F1dGhvcj48YXV0

aG9yPktpbSwgSi4gUy48L2F1dGhvcj48YXV0aG9yPkt3b24sIE0uIEguPC9hdXRob3I+PGF1dGhv

cj5LaW0sIFkuIFMuPC9hdXRob3I+PC9hdXRob3JzPjwvY29udHJpYnV0b3JzPjxhdXRoLWFkZHJl

c3M+RGVwYXJ0bWVudCBvZiBNb2xlY3VsYXIgU2NpZW5jZSBhbmQgVGVjaG5vbG9neSwgQWpvdSBV

bml2ZXJzaXR5LCBTdXdvbiA0NDMtNzQ5LCBLb3JlYS48L2F1dGgtYWRkcmVzcz48dGl0bGVzPjx0

aXRsZT5HZW5lIHNpbGVuY2luZyBieSBjZWxsLXBlbmV0cmF0aW5nLCBzZXF1ZW5jZS1zZWxlY3Rp

dmUgYW5kIG51Y2xlaWMtYWNpZCBoeWRyb2x5emluZyBhbnRpYm9kaWVzPC90aXRsZT48c2Vjb25k

YXJ5LXRpdGxlPk51Y2xlaWMgQWNpZHMgUmVzPC9zZWNvbmRhcnktdGl0bGU+PGFsdC10aXRsZT5O

dWNsZWljIGFjaWRzIHJlc2VhcmNoPC9hbHQtdGl0bGU+PC90aXRsZXM+PHBlcmlvZGljYWw+PGZ1

bGwtdGl0bGU+TnVjbGVpYyBBY2lkcyBSZXM8L2Z1bGwtdGl0bGU+PGFiYnItMT5OdWNsZWljIGFj

aWRzIHJlc2VhcmNoPC9hYmJyLTE+PC9wZXJpb2RpY2FsPjxhbHQtcGVyaW9kaWNhbD48ZnVsbC10

aXRsZT5OdWNsZWljIEFjaWRzIFJlczwvZnVsbC10aXRsZT48YWJici0xPk51Y2xlaWMgYWNpZHMg

cmVzZWFyY2g8L2FiYnItMT48L2FsdC1wZXJpb2RpY2FsPjxwYWdlcz4xNTk2LTYwOTwvcGFnZXM+

PHZvbHVtZT4zODwvdm9sdW1lPjxudW1iZXI+NTwvbnVtYmVyPjxlZGl0aW9uPjIwMDkvMTIvMTc8

L2VkaXRpb24+PGtleXdvcmRzPjxrZXl3b3JkPkFudGlib2RpZXMsIENhdGFseXRpYy9jaGVtaXN0

cnkvKm1ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+QXBvcHRvc2lzPC9rZXl3b3JkPjxrZXl3

b3JkPkJhc2UgU2VxdWVuY2U8L2tleXdvcmQ+PGtleXdvcmQ+Q2VsbCBMaW5lLCBUdW1vcjwva2V5

d29yZD48a2V5d29yZD5DeXRvc29sL21ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+RE5BLCBT

aW5nbGUtU3RyYW5kZWQvbWV0YWJvbGlzbTwva2V5d29yZD48a2V5d29yZD5HZW5lIEtub2NrZG93

biBUZWNobmlxdWVzPC9rZXl3b3JkPjxrZXl3b3JkPkdlbmUgTGlicmFyeTwva2V5d29yZD48a2V5

d29yZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdvcmQ+SHlkcm9seXNpczwva2V5d29yZD48a2V5d29y

ZD5JbW11bm9nbG9idWxpbiBMaWdodCBDaGFpbnMvY2hlbWlzdHJ5PC9rZXl3b3JkPjxrZXl3b3Jk

PkltbXVub2dsb2J1bGluIFZhcmlhYmxlIFJlZ2lvbi9jaGVtaXN0cnk8L2tleXdvcmQ+PGtleXdv

cmQ+KlJOQSBJbnRlcmZlcmVuY2U8L2tleXdvcmQ+PGtleXdvcmQ+Uk5BLCBNZXNzZW5nZXIvKm1l

dGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+UmVjZXB0b3IsIGVyYkItMi9hbnRhZ29uaXN0cyAm

YW1wOyBpbmhpYml0b3JzL2dlbmV0aWNzL21ldGFib2xpc208L2tleXdvcmQ+PC9rZXl3b3Jkcz48

ZGF0ZXM+PHllYXI+MjAxMDwveWVhcj48cHViLWRhdGVzPjxkYXRlPk1hcjwvZGF0ZT48L3B1Yi1k

YXRlcz48L2RhdGVzPjxpc2JuPjEzNjItNDk2MiAoRWxlY3Ryb25pYykmI3hEOzAzMDUtMTA0OCAo

TGlua2luZyk8L2lzYm4+PGFjY2Vzc2lvbi1udW0+MjAwMDc2MDI8L2FjY2Vzc2lvbi1udW0+PHdv

cmstdHlwZT5SZXNlYXJjaCBTdXBwb3J0LCBOb24tVS5TLiBHb3YmYXBvczt0PC93b3JrLXR5cGU+

PHVybHM+PHJlbGF0ZWQtdXJscz48dXJsPmh0dHA6Ly93d3cubmNiaS5ubG0ubmloLmdvdi9wdWJt

ZWQvMjAwMDc2MDI8L3VybD48L3JlbGF0ZWQtdXJscz48L3VybHM+PGN1c3RvbTI+MjgzNjU3Mjwv

Y3VzdG9tMj48ZWxlY3Ryb25pYy1yZXNvdXJjZS1udW0+MTAuMTA5My9uYXIvZ2twMTE0NTwvZWxl

Y3Ryb25pYy1yZXNvdXJjZS1udW0+PGxhbmd1YWdlPmVuZzwvbGFuZ3VhZ2U+PC9yZWNvcmQ+PC9D

aXRlPjwvRW5kTm90ZT4A

ADDIN EN.CITE.DATA 3, 6 Reactions were performed at 37 °C for 1 h and then terminated by incubating with 50 mM EDTA for 10 min at 37 °C. Samples were analyzed on 0.7 % agarose gel electrophoresis.Live cell imaging Live-cell time-lapse imaging was performed with Nikon A1 confocal system mounted on eclipse T1 microscope and using a 63× oil objective (NA 1.4). The cell incubation chamber was maintained with humidified atmosphere of 5 % CO2 and 37 °C. TMab4 labeled with Alexa Fluor 488 antibody labeling kit (Molecular Probes, Invitrogen) were excited at 488 nm and detected with a 500-530 nm band-pass filter, simultaneously acquiring differential-interference-contrast (DIC) images with excitation at 488 nm and detection in the transmission channel. For imaging, laser power was 0.5 % at 488 nm and gain value was 100. To quantify internalized TMab4 in cytosol for time course, cytosolic area of cells was set as region of interest (ROI) and mean fluorescence intensities were measured in all time points using ImageJ software. Fluorescence photobleaching was compensated by measuring the fluorescence of entire image and applying decreased fluorescence ratio. After background correction, all data were plotted using SigmaPlot software. Flow cytometryTo determine the internalization efficiency of cytotransmabs, HeLa cells were incubated with 1 μM of indicated antibodies for 6 h at 37 °C. After washing the cells with PBS and subsequently with the low pH glycine buffer to remove nonspecifically cell surface-bound antibodies, the cells were trypsinized, fixed, permeabilized, and then blocked prior to the staining of internalized cytotransmabs with Alexa Fluor 488-conjugated anti-human IgG antibody. About 1 × 104 of cells were analyzed using a FACS Calibur flow cytometer (Becton-Dickinson, Franklin Lakes, NJ), as previously described.PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5TdW5nPC9BdXRob3I+PFllYXI+MjAxMDwvWWVhcj48UmVj

TnVtPjYxPC9SZWNOdW0+PERpc3BsYXlUZXh0PjxzdHlsZSBmYWNlPSJzdXBlcnNjcmlwdCI+MTM8

L3N0eWxlPjwvRGlzcGxheVRleHQ+PHJlY29yZD48cmVjLW51bWJlcj42MTwvcmVjLW51bWJlcj48

Zm9yZWlnbi1rZXlzPjxrZXkgYXBwPSJFTiIgZGItaWQ9IjVzejBlNTJ6c3Nhd3hjZWEyeHBwdzU1

OGZyejV6enMyZDAydiI+NjE8L2tleT48L2ZvcmVpZ24ta2V5cz48cmVmLXR5cGUgbmFtZT0iSm91

cm5hbCBBcnRpY2xlIj4xNzwvcmVmLXR5cGU+PGNvbnRyaWJ1dG9ycz48YXV0aG9ycz48YXV0aG9y

PlN1bmcsIEUuIFMuPC9hdXRob3I+PGF1dGhvcj5LaW0sIEEuPC9hdXRob3I+PGF1dGhvcj5QYXJr

LCBKLiBTLjwvYXV0aG9yPjxhdXRob3I+Q2h1bmcsIEouPC9hdXRob3I+PGF1dGhvcj5Ld29uLCBN

LiBILjwvYXV0aG9yPjxhdXRob3I+S2ltLCBZLiBTLjwvYXV0aG9yPjwvYXV0aG9ycz48L2NvbnRy

aWJ1dG9ycz48YXV0aC1hZGRyZXNzPkRlcGFydG1lbnQgb2YgTW9sZWN1bGFyIFNjaWVuY2UgYW5k

IFRlY2hub2xvZ3ksIEFqb3UgVW5pdmVyc2l0eSwgU2FuNSwgV29uY2hlb24tZG9uZywgWWVvbmd0

b25nLWd1LCBTdXdvbiwgNDQzLTc0OSwgS29yZWEuPC9hdXRoLWFkZHJlc3M+PHRpdGxlcz48dGl0

bGU+SGlzdG9uZSBkZWFjZXR5bGFzZSBpbmhpYml0b3JzIHN5bmVyZ2lzdGljYWxseSBwb3RlbnRp

YXRlIGRlYXRoIHJlY2VwdG9yIDQtbWVkaWF0ZWQgYXBvcHRvdGljIGNlbGwgZGVhdGggb2YgaHVt

YW4gVC1jZWxsIGFjdXRlIGx5bXBob2JsYXN0aWMgbGV1a2VtaWEgY2VsbHM8L3RpdGxlPjxzZWNv

bmRhcnktdGl0bGU+QXBvcHRvc2lzPC9zZWNvbmRhcnktdGl0bGU+PC90aXRsZXM+PHBlcmlvZGlj

YWw+PGZ1bGwtdGl0bGU+QXBvcHRvc2lzPC9mdWxsLXRpdGxlPjwvcGVyaW9kaWNhbD48cGFnZXM+

MTI1Ni02OTwvcGFnZXM+PHZvbHVtZT4xNTwvdm9sdW1lPjxudW1iZXI+MTA8L251bWJlcj48ZWRp

dGlvbj4yMDEwLzA2LzI5PC9lZGl0aW9uPjxrZXl3b3Jkcz48a2V5d29yZD5BbnRpYm9kaWVzLCBN

b25vY2xvbmFsL3BoYXJtYWNvbG9neTwva2V5d29yZD48a2V5d29yZD5BbnRpbmVvcGxhc3RpYyBB

Z2VudHMvcGhhcm1hY29sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPkFwb3B0b3Npcy8qZHJ1ZyBlZmZl

Y3RzPC9rZXl3b3JkPjxrZXl3b3JkPkJsb3R0aW5nLCBXZXN0ZXJuPC9rZXl3b3JkPjxrZXl3b3Jk

PkNhc3Bhc2VzL21ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+Q2VsbCBMaW5lLCBUdW1vcjwv

a2V5d29yZD48a2V5d29yZD5Eb3duLVJlZ3VsYXRpb248L2tleXdvcmQ+PGtleXdvcmQ+RHJ1ZyBT

eW5lcmdpc208L2tleXdvcmQ+PGtleXdvcmQ+RmxvdyBDeXRvbWV0cnk8L2tleXdvcmQ+PGtleXdv

cmQ+SGlzdG9uZSBEZWFjZXR5bGFzZSBJbmhpYml0b3JzLypwaGFybWFjb2xvZ3k8L2tleXdvcmQ+

PGtleXdvcmQ+SGlzdG9uZSBEZWFjZXR5bGFzZXMvbWV0YWJvbGlzbTwva2V5d29yZD48a2V5d29y

ZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdvcmQ+SHlkcm94YW1pYyBBY2lkcy8qcGhhcm1hY29sb2d5

PC9rZXl3b3JkPjxrZXl3b3JkPkluaGliaXRvciBvZiBBcG9wdG9zaXMgUHJvdGVpbnMvbWV0YWJv

bGlzbTwva2V5d29yZD48a2V5d29yZD5NaXRvY2hvbmRyaWEvZHJ1ZyBlZmZlY3RzL21ldGFib2xp

c208L2tleXdvcmQ+PGtleXdvcmQ+UHJlY3Vyc29yIFQtQ2VsbCBMeW1waG9ibGFzdGljIExldWtl

bWlhLUx5bXBob21hL21ldGFib2xpc20vKnBhdGhvbG9neTwva2V5d29yZD48a2V5d29yZD5STkEs

IFNtYWxsIEludGVyZmVyaW5nPC9rZXl3b3JkPjxrZXl3b3JkPlJlY2VwdG9ycywgVHVtb3IgTmVj

cm9zaXMgRmFjdG9yLyppbW11bm9sb2d5LyptZXRhYm9saXNtPC9rZXl3b3JkPjxrZXl3b3JkPlNp

Z25hbCBUcmFuc2R1Y3Rpb24vZHJ1ZyBlZmZlY3RzPC9rZXl3b3JkPjxrZXl3b3JkPlRORi1SZWxh

dGVkIEFwb3B0b3Npcy1JbmR1Y2luZyBMaWdhbmQvbWV0YWJvbGlzbS9waGFybWFjb2xvZ3k8L2tl

eXdvcmQ+PGtleXdvcmQ+VmFscHJvaWMgQWNpZC8qcGhhcm1hY29sb2d5PC9rZXl3b3JkPjwva2V5

d29yZHM+PGRhdGVzPjx5ZWFyPjIwMTA8L3llYXI+PHB1Yi1kYXRlcz48ZGF0ZT5PY3Q8L2RhdGU+

PC9wdWItZGF0ZXM+PC9kYXRlcz48aXNibj4xNTczLTY3NVggKEVsZWN0cm9uaWMpJiN4RDsxMzYw

LTgxODUgKExpbmtpbmcpPC9pc2JuPjxhY2Nlc3Npb24tbnVtPjIwNTgyNDc3PC9hY2Nlc3Npb24t

bnVtPjx1cmxzPjxyZWxhdGVkLXVybHM+PHVybD5odHRwOi8vd3d3Lm5jYmkubmxtLm5paC5nb3Yv

cHVibWVkLzIwNTgyNDc3PC91cmw+PC9yZWxhdGVkLXVybHM+PC91cmxzPjxlbGVjdHJvbmljLXJl

c291cmNlLW51bT4xMC4xMDA3L3MxMDQ5NS0wMTAtMDUyMS05PC9lbGVjdHJvbmljLXJlc291cmNl

LW51bT48bGFuZ3VhZ2U+ZW5nPC9sYW5ndWFnZT48L3JlY29yZD48L0NpdGU+PC9FbmROb3RlPn==

ADDIN EN.CITE PEVuZE5vdGU+PENpdGU+PEF1dGhvcj5TdW5nPC9BdXRob3I+PFllYXI+MjAxMDwvWWVhcj48UmVj

TnVtPjYxPC9SZWNOdW0+PERpc3BsYXlUZXh0PjxzdHlsZSBmYWNlPSJzdXBlcnNjcmlwdCI+MTM8

L3N0eWxlPjwvRGlzcGxheVRleHQ+PHJlY29yZD48cmVjLW51bWJlcj42MTwvcmVjLW51bWJlcj48

Zm9yZWlnbi1rZXlzPjxrZXkgYXBwPSJFTiIgZGItaWQ9IjVzejBlNTJ6c3Nhd3hjZWEyeHBwdzU1

OGZyejV6enMyZDAydiI+NjE8L2tleT48L2ZvcmVpZ24ta2V5cz48cmVmLXR5cGUgbmFtZT0iSm91

cm5hbCBBcnRpY2xlIj4xNzwvcmVmLXR5cGU+PGNvbnRyaWJ1dG9ycz48YXV0aG9ycz48YXV0aG9y

PlN1bmcsIEUuIFMuPC9hdXRob3I+PGF1dGhvcj5LaW0sIEEuPC9hdXRob3I+PGF1dGhvcj5QYXJr

LCBKLiBTLjwvYXV0aG9yPjxhdXRob3I+Q2h1bmcsIEouPC9hdXRob3I+PGF1dGhvcj5Ld29uLCBN

LiBILjwvYXV0aG9yPjxhdXRob3I+S2ltLCBZLiBTLjwvYXV0aG9yPjwvYXV0aG9ycz48L2NvbnRy

aWJ1dG9ycz48YXV0aC1hZGRyZXNzPkRlcGFydG1lbnQgb2YgTW9sZWN1bGFyIFNjaWVuY2UgYW5k

IFRlY2hub2xvZ3ksIEFqb3UgVW5pdmVyc2l0eSwgU2FuNSwgV29uY2hlb24tZG9uZywgWWVvbmd0

b25nLWd1LCBTdXdvbiwgNDQzLTc0OSwgS29yZWEuPC9hdXRoLWFkZHJlc3M+PHRpdGxlcz48dGl0

bGU+SGlzdG9uZSBkZWFjZXR5bGFzZSBpbmhpYml0b3JzIHN5bmVyZ2lzdGljYWxseSBwb3RlbnRp

YXRlIGRlYXRoIHJlY2VwdG9yIDQtbWVkaWF0ZWQgYXBvcHRvdGljIGNlbGwgZGVhdGggb2YgaHVt

YW4gVC1jZWxsIGFjdXRlIGx5bXBob2JsYXN0aWMgbGV1a2VtaWEgY2VsbHM8L3RpdGxlPjxzZWNv

bmRhcnktdGl0bGU+QXBvcHRvc2lzPC9zZWNvbmRhcnktdGl0bGU+PC90aXRsZXM+PHBlcmlvZGlj

YWw+PGZ1bGwtdGl0bGU+QXBvcHRvc2lzPC9mdWxsLXRpdGxlPjwvcGVyaW9kaWNhbD48cGFnZXM+

MTI1Ni02OTwvcGFnZXM+PHZvbHVtZT4xNTwvdm9sdW1lPjxudW1iZXI+MTA8L251bWJlcj48ZWRp

dGlvbj4yMDEwLzA2LzI5PC9lZGl0aW9uPjxrZXl3b3Jkcz48a2V5d29yZD5BbnRpYm9kaWVzLCBN

b25vY2xvbmFsL3BoYXJtYWNvbG9neTwva2V5d29yZD48a2V5d29yZD5BbnRpbmVvcGxhc3RpYyBB

Z2VudHMvcGhhcm1hY29sb2d5PC9rZXl3b3JkPjxrZXl3b3JkPkFwb3B0b3Npcy8qZHJ1ZyBlZmZl

Y3RzPC9rZXl3b3JkPjxrZXl3b3JkPkJsb3R0aW5nLCBXZXN0ZXJuPC9rZXl3b3JkPjxrZXl3b3Jk

PkNhc3Bhc2VzL21ldGFib2xpc208L2tleXdvcmQ+PGtleXdvcmQ+Q2VsbCBMaW5lLCBUdW1vcjwv

a2V5d29yZD48a2V5d29yZD5Eb3duLVJlZ3VsYXRpb248L2tleXdvcmQ+PGtleXdvcmQ+RHJ1ZyBT

eW5lcmdpc208L2tleXdvcmQ+PGtleXdvcmQ+RmxvdyBDeXRvbWV0cnk8L2tleXdvcmQ+PGtleXdv

cmQ+SGlzdG9uZSBEZWFjZXR5bGFzZSBJbmhpYml0b3JzLypwaGFybWFjb2xvZ3k8L2tleXdvcmQ+

PGtleXdvcmQ+SGlzdG9uZSBEZWFjZXR5bGFzZXMvbWV0YWJvbGlzbTwva2V5d29yZD48a2V5d29y

ZD5IdW1hbnM8L2tleXdvcmQ+PGtleXdvcmQ+SHlkcm94YW1pYyBBY2lkcy8qcGhhcm1hY29sb2d5

PC9rZXl3b3JkPjxrZXl3b3JkPkluaGliaXRvciBvZiBBcG9wdG9zaXMgUHJvdGVpbnMvbWV0YWJv

bGlzbTwva2V5d29yZD48a2V5d29yZD5NaXRvY2hvbmRyaWEvZHJ1ZyBlZmZlY3RzL21ldGFib2xp

c208L2tleXdvcmQ+PGtleXdvcmQ+UHJlY3Vyc29yIFQtQ2VsbCBMeW1waG9ibGFzdGljIExldWtl

bWlhLUx5bXBob21hL21ldGFib2xpc20vKnBhdGhvbG9neTwva2V5d29yZD48a2V5d29yZD5STkEs

IFNtYWxsIEludGVyZmVyaW5nPC9rZXl3b3JkPjxrZXl3b3JkPlJlY2VwdG9ycywgVHVtb3IgTmVj

cm9zaXMgRmFjdG9yLyppbW11bm9sb2d5LyptZXRhYm9saXNtPC9rZXl3b3JkPjxrZXl3b3JkPlNp

Z25hbCBUcmFuc2R1Y3Rpb24vZHJ1ZyBlZmZlY3RzPC9rZXl3b3JkPjxrZXl3b3JkPlRORi1SZWxh

dGVkIEFwb3B0b3Npcy1JbmR1Y2luZyBMaWdhbmQvbWV0YWJvbGlzbS9waGFybWFjb2xvZ3k8L2tl

eXdvcmQ+PGtleXdvcmQ+VmFscHJvaWMgQWNpZC8qcGhhcm1hY29sb2d5PC9rZXl3b3JkPjwva2V5

d29yZHM+PGRhdGVzPjx5ZWFyPjIwMTA8L3llYXI+PHB1Yi1kYXRlcz48ZGF0ZT5PY3Q8L2RhdGU+

PC9wdWItZGF0ZXM+PC9kYXRlcz48aXNibj4xNTczLTY3NVggKEVsZWN0cm9uaWMpJiN4RDsxMzYw

LTgxODUgKExpbmtpbmcpPC9pc2JuPjxhY2Nlc3Npb24tbnVtPjIwNTgyNDc3PC9hY2Nlc3Npb24t

bnVtPjx1cmxzPjxyZWxhdGVkLXVybHM+PHVybD5odHRwOi8vd3d3Lm5jYmkubmxtLm5paC5nb3Yv

cHVibWVkLzIwNTgyNDc3PC91cmw+PC9yZWxhdGVkLXVybHM+PC91cmxzPjxlbGVjdHJvbmljLXJl

c291cmNlLW51bT4xMC4xMDA3L3MxMDQ5NS0wMTAtMDUyMS05PC9lbGVjdHJvbmljLXJlc291cmNl

LW51bT48bGFuZ3VhZ2U+ZW5nPC9sYW5ndWFnZT48L3JlY29yZD48L0NpdGU+PC9FbmROb3RlPn==

ADDIN EN.CITE.DATA 13Supplementary References ADDIN EN.REFLIST 1.Kabat EA, Wu TT. Identical V region amino acid sequences and segments of sequences in antibodies of different specificities. Relative contributions of VH and VL genes, minigenes, and complementarity-determining regions to binding of antibody-combining sites. J Immunol 1991; 147:1709-19.2.Jang JY, Jeong JG, Jun HR, Lee SC, Kim JS, Kim YS, et al. A nucleic acid-hydrolyzing antibody penetrates into cells via caveolae-mediated endocytosis, localizes in the cytosol and exhibits cytotoxicity. Cell Mol Life Sci 2009; 66:1985-97.3.Lee WR, Jang JY, Kim JS, Kwon MH, Kim YS. Gene silencing by cell-penetrating, sequence-selective and nucleic-acid hydrolyzing antibodies. Nucleic Acids Res 2010; 38:1596-609.4.Hu S, Liang S, Guo H, Zhang D, Li H, Wang X, et al. Comparison of the inhibition mechanisms of adalimumab and infliximab in treating tumor necrosis factor alpha-associated diseases from a molecular view. J Biol Chem 2013; 288:27059-67.5.Vargas-Madrazo E, Paz-Garcia E. An improved model of association for VH-VL immunoglobulin domains: asymmetries between VH and VL in the packing of some interface residues. J Mol Recognit 2003; 16:113-20.6.Kim DS, Lee SH, Kim JS, Lee SC, Kwon MH, Kim YS. Generation of humanized anti-DNA hydrolyzing catalytic antibodies by complementarity determining region grafting. Biochem Biophys Res Commun 2009; 379:314-8.7.Kim DG, Lee JY, Kwon NH, Fang P, Zhang Q, Wang J, et al. Chemical inhibition of prometastatic lysyl-tRNA synthetase-laminin receptor interaction. Nat Chem Biol 2014; 10:29-34.8.Lee SH, Park DW, Sung ES, Park HR, Kim JK, Kim YS. Humanization of an agonistic anti-death receptor 4 single chain variable fragment antibody and avidity-mediated enhancement of its cell death-inducing activity. Mol Immunol 2010; 47:816-24.9.Shin TH, Sung ES, Kim YJ, Kim KS, Kim SH, Kim SK, et al. Enhancement of the tumor penetration of monoclonal antibody by fusion of a neuropilin-targeting Peptide improves the antitumor efficacy. Mol Cancer Ther 2014; 13:651-61.10.Lee YH, Iijima M, Kado Y, Mizohata E, Inoue T, Sugiyama A, et al. Construction and characterization of functional anti-epiregulin humanized monoclonal antibodies. Biochem Biophys Res Commun 2013; 441:1011-7.11.Pericleous LM, Richards J, Epenetos AA, Courtenay-Luck N, Deonarain MP. Characterisation and internalisation of recombinant humanised HMFG-1 antibodies against MUC1. Br J Cancer 2005; 93:1257-66.12.Troise F, Cafaro V, Giancola C, D'Alessio G, De Lorenzo C. Differential binding of human immunoagents and Herceptin to the ErbB2 receptor. FEBS J 2008; 275:4967-79.13.Sung ES, Kim A, Park JS, Chung J, Kwon MH, Kim YS. Histone deacetylase inhibitors synergistically potentiate death receptor 4-mediated apoptotic cell death of human T-cell acute lymphoblastic leukemia cells. Apoptosis 2010; 15:1256-69. ................
................

In order to avoid copyright disputes, this page is only a partial summary.

Google Online Preview   Download