Ion exchange chromatography columns and resins
Ion exchange
chromatography
columns and resins
Selection guide
Introduction to ion exchange
chromatography
What is ion exchange (IEX) chromatography?
IEX is a liquid chromatography technique to separate proteins that have only slight differences in
their net surface charge. Even very closely related proteins will have some difference in charge and
can be effectively separated using this purification method.
The chromatography technique is based on the interaction of a charged molecule
and the oppositely charged chromatography resin.
This chromatography technique takes advantage of the fact that the relationship
between net surface charge and pH is unique for a specific protein.
Positively charged protein
(blue) binds to negatively
charged resin beads (gray)
Typically, conditions are selected to ensure that the molecules of interest bind to
the resin as they are loaded onto the column.
Conditions are then altered so that the bound substances are eluted differently.
Negatively charged protein
(orange) flows through
IEX is performed in four main steps as shown below.
Fig 1. Ion exchange chromatography using a
cation exchanger.
How does ion exchange chromatography work?
1. Equilibration
2. Sample application and wash
Prepare the column
to the desired start
conditions.
3. Elution
4. Regeneration
Biomolecules are
gradually released
from the ionic
exchanger by a
change in the
buffer composition.
Fig 2. Main steps of an ion exchange chromatography run.
2
Bind the target
molecules and wash
out all unbound
material.
Removal of all
molecules still bound.
Select your ion exchange
chromatography resin
Select the chromatography resin according to the objective of the purification step and the condition
of the starting material. Other factors such as sample stability, scale, speed, binding capacity, and
S
equipment available may also influence the final choice. Use Cthe
trees on pages 4 and 5 to
apto decision
find the most appropriate resin for your needs.
Q
Capto
Cation r1
ge
exchan
Capto
Strongnger1
ha
ion exc
Anion r
ge
exchan
re
Captu
Capto
IEX
Capto
P
pAct
p.
S Im5
Capto
Screen t
n
differe s
in
IEX res
mAb n
tio
separa
Capto
Anion r
ge
exchan
Res
SP Imp
Capto
Cation r
ge
exchan
es
Q ImpR
Capto
mAb n
tio
separa
lute
Bind/ede
mo
Cation r
ge
exchan
hrough
Flow-tode1
m
lute
Bind/ede
mo
ing
n kit
selectio
Resins for processning plates
ee
ing Scr
Polish
development
o
t
p
a
or C
reDict
p. 4
Polish
plates
Pre
select
(when tisfactory
is unsaCapto Q)
with
Screen t
n
differe s
in
IEX res
ediate
Intermcation
purifi
DEAE
ening
? Scre
Dictor
ion
Weak ger
exchan ivity
Resins for
research use
ss
Procem
p ent
develo
Q XP
Anion r
ge
exchan
hrough
Flow-tode1
m
ct
S ImpA
Capto
Res
SP Imp
Capto
S
Capto
es
Q ImpR
Capto
Q
PreDic
ning
g Scree
olishin
plates
pto P
tor Ca
Screen t
n
differe s
in
IEX res
Ion exchange chromatography handbook
Looking for protocols and tips for using ion exchange
chromatography?
Download our handbook from handbooks
3
Guide to IEX resins for research
applications1
Highest
resolution2
(e.g., structural
biology and
protein analysis)
550
Capto? HiRes columns
Q (anion exchanger)
S (cation exchanger)
500
450
A280 (mAU)
400
A280 (mAU)
A280 (mAU)
Small-scale
preparative
/analytical
350
550
300
500
250
450
200
400
150
350
100
550
300
50
500
250
0
450
200
400
150
350
100
550
300
550
50
500
250
500
0
450
200
450
400
150
400
350
100
350
300
550
50
300
250
500
0
250
200
450
200
150
400
150
100
350
100
550
50
300
50
500
0
250
0
450
200
400
150
350
100
550
300
550
50
500
250
500
0
450
200
450
400
150
400
350
100
350
300
550
50
300
250
500
0
250
200
450
200
150
400
150
100
350
100
550
50
300
50
500
0
250
0
450
200
400
150
350
100
550
300
550
50
500
250
500
0
450
200
450
400
150
400
350
100
350
300
50
550
300
250
0
500
250
200
450
200
150
400
150
100
350
100
50
550
300
50
0
500
250
0
450
200
400
150
350
100
550
300
50
500
250
0
450
200
400
150
350
100
300
50
250
0
200
6
8
10
12
14
16
18
20
22
24
26
28
30
Volume (mL)
Separation3 on a Capto HiRes Q 5/50 column
SOURCE? 15
High resolution
at fast flow rates
20
22
24
26
28
30
20
22
24
26
28
30
16 18 20
16 18 20
Volume (mL)
Volume (mL)
22
22
24
24
26
26
28
28
30
30
22
24
26
28
30
Separation3 on a RESOURCE? Q 1mL
column
(SOURCE
15 resin)
6
8
10 12 14 16 18 20 22 24 26
28
30
A280
A280
(mAU)
(mAU)
Research
use
Preparative
purification
Sepharose? High Performance
Routine
purification
A280A(mAU)
(mAU)
280
A280 (mAU)
Q (anion exchanger)
S (cation exchanger)
A280 (mAU)
A280
A280
(mAU)
(mAU)
A280 (mAU)
A280 (mAU)
Q (anion exchanger)
S (cation exchanger)
6
8
10
12
14
16
18
Volume (mL)
6
8
10
12
14
16
18
Volume (mL)
6
6
8
8
10
10
12
12
14
14
6
8
10
12
14
16
18
20
Volume (mL)
Volume (mL)
6
6
8
8
10
10
12
12
14
14
6
8
10
12
14
16 18 20
16 18 20
Volume (mL)
Volume (mL)
16
18
22
22
24
24
26
26
28
28
30
30
20
22
24
26
28
30
20
22
24
26
28
30
Volume (mL)
6
8
10
12
14
16
18
Volume (mL)
Routine purification
with faster loading
of viscous samples
(e.g., E. coli lysates)
A280 (mAU)
A280 (mAU)
Separation3 on a HiTrap Q HP 1mL column
(Sepharose
High Performance resin)
6
8
10 12 14 16 18 20 22 24 26 28 30
Capto ImpRes
A280 (mAU)
Q (anion exchanger)
S (cation exchanger)
PP
H
G
H
S
PP
PP
G
96
G
S
150
L
S
96
96
L
6
8
10
12
14 Volume
16 (mL)
18 20
22
24
26
28
30
20
22
24
26
28
30
20
22
24
26
28
30
20
22
24
26
28
30
Volume (mL)
6
8
10
12
14
16
18
Volume (mL)
6
8
10
12
14
16
18
Volume (mL)
100
L
50
0
6
8
10
12
14
16
18
Volume (mL)
Separation on a HiTrap Capto Q ImpRes
1 mL column
3
Format guide
Type of purification
System purification
Bottles of
chromatography resins
Small-column cartridges
(1 and 5 mL)
Other columns
Format name
Lab pack
HiTrap?
HiScreen?, HiPrep?,
RESOURCE, Tricorn?
Use
4
Manual or system purification
Format
Batch purification
and packing empty
columns
Easy to use with a
syringe, peristaltic pump,
or a chromatography
system
Larger scale or highperformance applications
1
This resin selection is our recommendation for the most common
needs in research. For a complete list of products, refer to the
ordering information table.
2
Compared with other IEX columns from GE Healthcare Life Sciences.
3
Protein mix purified at 1 mL/min and containing:
Apotransferrin
¦Á¨CLactalalbumin
¦Â-Lactoglobulin
Amyloglucosidase
Guide to IEX resins for process
development
Cation
exchanger1
Capto S
Capto Q
Strong
ion exchanger1
Capto Q XP
Anion
exchanger
Capture
Weak ion
exchanger
Capto DEAE
(when selectivity
is unsatisfactory
with Capto Q)
PreDictor? Screening plates
Screen
different
IEX resins
Capto IEX selection kit
Screen
different
IEX resins
PreDictor Capto Polishing Screening plates
mAb
separation
Process
development
Capto S ImpAct
Cation
exchanger
Intermediate
purification
Capto SP ImpRes
Anion
exchanger
Capto Q ImpRes
mAb
separation
Capto S ImpAct
Bind/elute
mode
Capto SP ImpRes
Cation
exchanger
Flow-through
mode1
H
Bind/elute
mode
Anion
exchanger
Polishing
PP
H
H
PP
G
Screen
H
H
different
IEX resins
PP
Capto S
Capto Q ImpRes
Flow-through
mode1
PP
G
G
S
G
PP
S
Capto Q
S
96
S
G
96
96
L
S
L
96
L
96
PreDictor Capto Polishing Screening plates
L
L
Format guide
1
2
Format
96-well filter
plates
(2, 6, 20, or 50 ?L)
Miniaturized
prepacked
columns (50,
200, and 600 ?L)
Bottles of
chromatography
resins
Prepacked
columns
(1 and 5 mL)
Prepacked
columns
(4.7 mL with
10 cm bed
height)
Single-use,
prepacked
columns
¡Ý1L
Format name
PreDictor
PreDictor
RoboColumn?
Bulk pack
HiTrap
HiScreen
ReadyToProcess?2
Application
High-throughput
screening
High-throughput
process
development
Batch purification
and self-packing
Process
development,
screening
Method
optimization
Clinical
manufacturing
We also offer solutions based on membrane adsorbers technology. Contact your sales representative.
For more information about ReadyToProcess columns, contact your sales representative.
5
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