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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