Characteristics of Multimodal Chromatography Media

Appendix 1, Extracted from Multimodal Chromatography (PDF), GE Healthcare, 2013

Table A1.1. Characteristics of Capto adhere

Matrix Highly Cross-linked Agarose
Functional group multimodal strong anion exchanger
Particle size (d50v)1 75 μm
Total ionic capacity 0.09 to 0.12 mmol Cl/mL medium
Flow velocity2 at least 600 cm/h in a 1 m diameter column with 20 cm bed height at 20°C using process buffers with the same viscosity as water at < 3 bar (0.3 MPa)
pH stability3
-working range
-cleaning-in-place (CIP)
3 to 12
2 to 14
Working temperature4 4°C to 30°C
Chemical stability5 all commonly used aqueous buffers, 1 M acetic acid, 1 M sodium hydroxide
Avoid oxidizing agents, anionic detergents

1 d50v is the median particle size of the cumulative volume distribution.

2 The capacity for selective removal of some key contaminants may decrease at high flow velocity

3 Working range: pH interval where the medium can be operated without significant change in function.

CIP: pH stability where the medium can be subjected to cleaning- or sanitization-in-place without significant change in function.

4 Capto adhere can be used under cold-room conditions, but the capacity for some key contaminants may decrease.

5 No significant change in ionic binding capacity and carbon content after 1 wk storage in 1M NaOH at 40°C.

Table A1.2. Characteristics of Capto adhere ImpRes

Matrix Highly Cross-linked Agarose
Functional group multimodal strong anion exchanger
Particle size (d50v)1 36 to 44 μm
Total ionic capacity 0.08 to 0.11 mmol Cl/mL medium
Flow velocity2 at least 220 cm/h in a 1 m diameter column with 20 cm bed height at 20°C using process buffers with the same viscosity as water at < 3 bar (0.3 MPa)
pH stability3
-working range
-CIP
3 to 12
2 to 14
Working temperature4 4°C to 30°C
Chemical stability5 all commonly used aqueous buffers, 1 M acetic acid, 1 M sodium hydroxide
Storage 4°C to 30°C in 20% ethanol
Regulatory support Regulatory support file is available

1 d50v is the average particle size of the cumulative volume distribution.

2 The capacity for selective removal of some key contaminants may decrease at high flow velocity.

3 Working range: pH interval where the medium can be operated without significant change in function.

CIP: pH stability where the medium can be subjected to CIP without significant change in function.

4 Capto adhere ImpRes can be used under cold-room conditions, but the capacity for some key contaminants may decrease.

5 No significant change in nitrogen and carbon content after 1 wk storage in 1 M NaOH at 40°C.

Table A1.3. Characteristics of Capto MMC

Matrix highly cross-linked agarose
Functional group multimodal weak cation exchanger
Particle size (d50v)1 75 μm
Total ionic capacity 0.07 to 0.09 mmol Cl/mL medium
Flow velocity2 at least 600 cm/h in a 1 m diameter column with 20 cm bed height at 20°C using process buffers with the same viscosity as water at < 3 bar (0.3 MPa)
DBC2 > 45 mg BSA/mL medium at 30 mS/cm
pH stability3
-short term
-long term
2 to 14
2 to 12
Working temperature4 4°C to 30°C
Chemical stability5 all commonly used aqueous buffers, 1 M acetic acid, 1 M sodium hydroxide5, 8 M urea, 6 M guanidine hydrochloride, and 70% ethanol
Avoid oxidizing agents, cationic detergents

1 d50v is the median particle size of the cumulative volume distribution.

2 DBC at 10% breakthrough as measured at a residence time of 2 min, 300 cm/h in a Tricorn 5/100 column with 10 cm bed height in 50 mM Na-acetate, pH 4.75, 250 mM NaCl.

3 Short term pH: pH interval that the medium can be subjected to, for cleaning- or sanitization-in-place (accumulated 90 to 300 h at room temperature) without significant change in function.

Long term pH: pH interval where the medium can be operated without significant change in function.

4 Capto MMC can be used under cold-room conditions, but for some proteins the capacity may decrease.

5 No significant change in ionic binding capacity and carbon content after 1 wk storage in 1 M NaOH at 40°C.

Table A1.4. Characteristics of Capto MMC ImpRes

Matrix Highly Cross-linked Agarose
Functional group multimodal weak cation exchanger
Particle size (d50v)1 36 to 44 μm
Ligand density 25 to 39 μmol ligand/mL medium
Maximum flow velocity2 at least 220 cm/h in a 1 m diameter column with 20 cm bed height at 20°C using process buffers with the same viscosity as water at < 3 bar (0.3 MPa)
pH stability3
-working range
-CIP
3 to 12
2 to 14
Working temperature4 4°C to 30°C
Chemical stability5 all commonly used aqueous buffers, 1 M acetic acid, 1 M sodium hydroxide
Storage 4°C to 30°C in 20% ethanol, 0.2 M sodium acetate
Regulatory support Regulatory support file is available

1 d50v is the average particle size of the cumulative volume distribution.

2 The capacity for selective removal of some key contaminants may decrease at high flow velocity.

3 Working range: pH interval where the medium can be operated without significant change in function.

CIP: pH stability where the medium can be subjected to CIP without significant change in function.

4 Capto MMC ImpRes can be used under cold-room conditions, but the capacity for some key contaminants may decrease.

5 No significant change in nitrogen, sulfur, and carbon content after 1 wk storage in 1 M NaOH at 40°C.

Table A1.5. Characteristics of Capto Core 700

Matrix Highly Flow Agarose
Size cut-off of outer layer Mr ≈ 700 000
pKa of protonated octylamine1 10.65
Functional group in the core CH3CH2CH2CH2CH2CH2CH2CH2NH–
Ionic capacity/mL 40 to 85 μmol Cl/mL medium
Particle size (d50v)2 85 μm
Maximum operational flow velocity3 500 cm/h in columns with 20 cm bed height at < 2 bar (0.2 MPa)
Binding capacity4/mL 13 mg ovalbumin/mL medium
pH stability
-CIP
-working range
2 to 14
3 to 13
Working temperature 4°C to 30°C
Chemical stability5 all commonly used aqueous buffers, 1 M sodium hydroxide5, 6 M guanidine hydrochloride, 30% isopropyl alcohol, and 70% ethanol
Avoid oxidizing agents, anionic detergents
Storage 20% ethanol

1 pKa of protonated octylamine before attachment to the medium. After attachment the pKa may be slightly different.

2 d50v is the average particle size of the cumulative volume distribution.

3 Maximum flow velocity that has been verified for long-term use.

4 DBC was measured at 10% breakthrough with a residence time of 3 min (1.6 mL/min = 200 cm/h) in HiScreen columns. The mobile phase was 20 mM Tris/HCl with 0.15 M NaCl at pH 7.5.

5 No significant change in ionic capacity and carbon content after 1 wk storage in 1 M NaOH at 40°C.

Materials

     
Related Links