Ion-exchange Chromatography Resins

Ion-exchange chromatography (IEX) is one of the most widely used chromatography techniques, which plays an important role in the separation and purification of biomolecules since the 1960s. IEC controls the reversible interaction between charged molecules and ion exchange media with opposite charges to achieve the binding and elution of specific molecules to achieve the separation effect. IEC widely uses in protein, skin, nucleic acids and other electric charges in the separation and purification of biomolecules. The advantages of high capacity and high resolution to well thousand among the whole process of capture, purification, and fine purification stage. IEC applies to the analysis of a small amount of sample, and the purification of sample preparation.

GALAK Ion-exchange Chromatography resins use agarose and monodisperse PS-DVB microspheres for substrates. For PS-DVB microspheres substrate, it has the advantages of high mechanical strength and excellent pressure resistance to meet the high requirements of preparative production conditions.

ion-exchange chromatography

GALAK Technology Advantages

GALAK ion-exchange packing materials are a series of ion exchange media developed based on monodisperse PSDVB microspheres. Its matrix structure and hydrophilic polysaccharide base material, such as agarose, cellulose, is completely different. First of all, it has excellent mechanical properties, can withstand up to 10MPa pressure; Secondly, the large pore size of 1000A can guarantee the free entry and exit of biological macromolecules. Finally, the special coating technology of GALAK gives it enough hydrophilicity to ensure its excellent biocompatibility. The monodisperse particle size can effectively reduce the column pressure and mass transfer obstruction

GALAK ion-exchange packing materials use "tentacles" surface derived technology, functional groups in the form of linear polymer chain covalently combined on the surface of the substrate. Relative to the traditional ion exchange medium directly on the surface modification or with only short-chain in combination with the pattern of functional groups, GALAK ion-exchange packing materials has more effective number of functional groups, not only its space steric hindrance to reduce macromolecular protein, antibody, viruses, and plasmid can more effectively combine with medium of functional groups, significantly increase capacity, and "tentacles" structure can effectively reduce enough biological molecules and nonspecific interaction between the dielectric substrate, so as to improve the recovery rate of target molecules.

galak-ion-exchange

GALAK Ion-exchange

traditional-ion-exchange

Traditional Ion-exchange

Sepromax PSDVB Resin

Sepromax® S50

Sepromax® CM50Sepromax® Q50

Sepromax® D50

Substrate

Rigid, PS-DVB microspheres

Particle Size

35-85 μm

Ligand

-SO3--COO--N+(CH3)3

-N+H(CH3)2

pH Range

2-126-122-12

2-9

pKa

14.513

8-9

Capacity

60mg hIgG/ml80mg Lysozyme/ml80mg Lysozyme/ml

100mg BSA/ml

Max Pressure

1500 psi (100 bar or 10 MPa)

pH Stability

1-14
Storage

20% EtOH,4-30℃

GLKgel Agarose Resin

GLKgel Strong Cation IEX SP Resin

SP 6BBSP 6FFSP 6HFSP 6HPSP 6XL

SP HPR

Substrate

6% cross-linked AgaroseHigh-rigid Agarose6% cross-linked Agarose6% cross-linked Agarose with glucan

High-rigid Agarose

Particle Size

200μm

(165-300μm)

90μm

(45-165μm)

90μm

(45-165μm)

37μm

(25-45μm)

90μm

(45-165μm)

37μm

(25-45μm)

Ligand

-CH2CH2CH2SO3-

Loading

Capacity

180-250μmol

H+/ml resin

140-200μmol H+/ml resin150-200μmol H+/ml resin180-250μmol H+/ml resin

130-160μmol H+/ml resin

pH Stability

4-13 (Long)

3-14 (Short)

4-12 (Long)

3-14 (Short)

4-13 (Long)

3-14 (Short)

4-12 (Long)

3-14 (Short)

Pressure

≤0.3MPa

≤0.5MPa

Flow Rate

1800cm/h700 cm/h100 cm/h150 cm/h700 cm/h

400 cm/h

Chemical Stability

All common buffer, 1.0m sodium hydroxide, 8.0m urea, 6.0m GuHCl, 70% ethanol

Avoid using oxidant, cationic detergent, cationic buffer

Storage

0.2M NaAc, 20% EtOH, 4-30℃

GLKgel Strong Anion IEX Q Resin

Q 6BBQ 6FFQ 6HFQ 6HPQ 6XLQ HPR
Substrate6% cross-linked AgaroseHigh-rigid Agarose6% cross-linked Agarose6% cross-linked Agarose with glucan

High-rigid Agarose

Particle Size

200μm

(165-300μm)

90μm

(45-165μm)

90μm

(45-165μm)

37μm

(25-45μm)

90μm

(45-165μm)

37μm

(25-45μm)

Ligand

-N+(CH3)3

Loading Capacity

180-250μmol

Cl-/ml resin

160-200μmol

Cl-/ml resin

140-200μmol

Cl-/ml resin

180-250μmol Cl-/ml resin150-180μmol Cl-/ml resin
pH Stability2-12 (Long Period)

2-14 (Short Period)

2-12 (Long)

2-14 (Short )

2-12 (Long)

2-14 (Short)

2-12 (Long )

2-14 (Short)

Pressure

≤0.3MPa≤0.5MPa
Flow Rate1800cm/h700 cm/h1000 cm/h150 cm/h700 cm/h

400 cm/h

Chemical Stability

All common buffer, 1.0m sodium hydroxide, 8.0m urea, 6.0m GuHCl, 70% ethanol

Avoid using oxidant, cationic detergent, cationic buffer

Storage

20% EtOH, 4-30℃

GLKgel Strong Anion IEX MMA Resin

SubstrateParticle SizeCapacitypH StabilityPressure

Flow Rate

MMA 6HF

High Rigid

Agarose

90μm

(45-165μm)

90-120μmol

Cl-/ml resin

2-14 (Long)

4-12 (Short)

≤0.5 MPa1000 cm/h
MMA HPRHigh Rigid

Agarose

37μm

(25-45μm)

80-110μmol

Cl-/ml resin

2-14 (Long)

4-12 (Short)

≤0.5 MPa

400 cm/h

Chemical Stability

All common buffer, 1.0m NaOH, 8.0m urea, 6.0m GuHCl, 70% ethanol

Avoid using oxidant, cationic detergent, cationic buffer

Storage

  20% EtOH, 4-30℃

GLKgel Weak Cation IEX CM Resin

 

CM 6FFCM 6HFCM 6HPCM 6XL
Substrate6% cross-linked

Agarose

High-rigid

Agarose

6% cross-linked

Agarose

6% cross-linked Agarose with glucan

Particle Size

90μm (45-165μm)90μm (45-165μm)37μm (25-45μm)90μm (45-165μm)
Ligand

-O-CH2COO-

Capacity

90-130μmol

H+/ml resin

90-120μmol

H+/ml resin

80-110μmol

H+/ml resin

180-250μmol

H+/ml resin

pH Stability4-13 (Long)

2-14 (Short)

4-12 (Long)

3-14 (Short)

4-13 (Long)

2-14 (Short)

Pressure

≤0.3 MPa≤0.5MPa≤0.3 MPa
Flow Rate700 cm/h1000 cm/h150 cm/h700 cm/h
Chemical

Stability

All common buffer, 1.0m NaOH, 8.0m urea, 6.0m guanidine hydrochloride, 70% ethanol

Avoid using oxidant, cationic detergent, cationic buffer

Storage

 20% EtOH, 4-30℃

GLKgel Weak Cation IEX MMC Resin

 

SubstrateParticle SizeCapacitypH StabilitypH StabilityFlow Rate
MMC 6HFHigh Rigid

Agarose

90μm

(45-165μm)

70-90μmol

H+/ml resin

2-14 (Long)

3-12 (Short)

≤0.5 MPa

1000 cm/h

MMC HPR

High Rigid

Agarose

37μm

(25-45μm)

60-80μmol

H+/ml resin

2-14 (Long)

3-12 (Short)

≤0.5 MPa400 cm/h

Chemical Stability

All common buffer, 1.0m NaOH, 8.0m urea, 6.0m guanidine hydrochloride, 70% ethanol

Avoid using oxidant, cationic detergent, cationic buffer

Storage

  20% EtOH, 4-30d

GLKgel Weak Anion IEX DEDA Resin

 

DEDA 6FFDEDA 6HFDEDA 6HP

DEDA 6XL

Substrate

6% cross-linked AgaroseHigh-rigid Agarose6% cross-linked Agarose6% cross-linked Agarose
Particle Size90μm (45-165μm)90μm (45-165μm)37μm (25-45μm)

90μm (45-165μm)

Ligand

-N+(CH3)3
Capacity110-160μmol

Cl-/ml resin

290-350μmol

Cl-/ml resin

90-130μmol

Cl-/ml resin

110-160μmol

Cl-/ml resin

pH Stability

2-13 (Long)

1-14 (Short)

2-12 (Long)

2-14 (Short)

2-13 (Long)

1-14 (Short)

Pressure≤0.3 MPa≤0.5MPa

≤0.3 MPa

Flow Rate

700 cm/h1000 cm/h150 cm/h700 cm/h
Chemical

Stability

 All common buffer, 1.0m NaOH, 8.0m urea, 6.0m guanidine hydrochloride, 70% ethanol

Avoid using oxidant, cationic detergent, cationic buffer

Storage

  20% EtOH, 4-30℃

GLKgel Weak Anion IEX ANX Resin

 

SubstrateParticle SizeLigandCapacitypH StabilitypH Stability

Flow Rate

ANX 4FF4% cross-linked Agarose90μm

(45-165μm)

-N+(C2H5)2H130-170μmol

Cl-/ml resin

3-10 (Long)

2-14 (Short)

≤0.3 MPa

250 cm/h

Chemical Stability

All common buffer, 1.0m NaOH, 8.0m urea, 6.0m guanidine hydrochloride, 70% ethanol

Avoid using oxidant, cationic detergent, cationic buffer

Storage

  20% EtOH, 4-30℃

Selection Of Ion-exchange Packing Materials

In practical applications, the substrate type, pore structure, particle size and distribution, substrate type, and density of ion-exchange media will affect the tomographic effect of the medium.

  • The porosity of the substrate provides a large surface area covered with charged groups, thus ensuring a high bonding capacity. However, the large pore size ensures the effective mass transfer of protein biomolecules.
  • The hydrophilic substrate has better biocompatibility and less non-specific adsorption.
  • The high chemical stability ensures that the medium can be cleaned with a vigorous cleaning solution if necessary.
  • The high physical stability ensures that the volume of the column bed. It remains constant when the concentration of salt ions or the pH value changes dramatically. This can increase the repeatability of the system, less unnecessary reloading columns. At the same time, the consistency of particle size distribution is beneficial to the operation of high-speed liquid flow, especially in cleaning or rebalancing steps, which can increase the flux and working capacity of the system.
Cation-exchange
Sulfonic Group S/SPStrong Type-SO32-
Carboxymethyl CMWeek Type-COO
Anion-exchange
Quaternary Ammonium  QStrong Type-N+(CH3)3
Tertiary Amine DWeek Type-N+H(CH3)2

Strong ion-exchange media have the following advantages

  • Since the charged nature of the medium does not change with the change of pH, the establishment and optimization of the separation process will be very fast and easy.
  • Since there is no intermediate form of charge interaction, the interaction mechanism is simple.
  • At high or low pH, the binding capacity of the sample is maintained because the ion-exchange medium does not lose charge.

Most proteins have isoelectric points between 5.5 and 7.5 and can be separated by a strong or weak ion exchange medium. The advantage of weak ion-exchange media, such as D and CM, is that they offer different selectivity than strong ion-exchange media. The disadvantage is that weak ion-exchange media gain or loses protons as pH changes, so their ion-exchange capacity changes as pH changes.

When the desired selectivity cannot be obtained with strong ion-exchange media (Q, S, SP), weak ion-exchange media such as D, CM can be tried. When using a weak ion exchanger, use a float that minimizes the effect in the following pH range:

  • D: pH 2-9
  • CM: pH 6-10

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