Columns

BioPro IEX Column

BioPro IEX Columns are for the separation of proteins, peptides, and nucleic acids. BioPro IEX Columns are available in QA and SP chemistries and are based on porous (QA and SP columns) and non-porous (QF and SF columns) hydrophilic polymer beads with low nonspecific adsorption. The porous materials offer excellent binding capacity with exceptionally high efficiency. The non-porous particles offer high efficiency and exceptional resolution.

Specification

  BioPro IEX QA
BioPro IEX SP
BioPro IEX QF BioPro IEX SF
Matrix hydrophilic porous polymer beads hydrophilic non-porous polymer beads
Particle size (µm) 5 3, 5
Charged group -CH2N+(CH3)3 -CH2CH2CH2SO3- -CH2N+(CH3)3 -CH2CH2CH2SO3-
Counter ion Cl- Na+ Cl- Na+
Ion-exchange capacity [reference value]
(meq/mL-resin)
0.09 0.09 0.09 0.24
Dynamic binding capacity [reference value]
(mg/mL-resin)
>110 (BSA) >70 (human-IgG) >12 (BSA) >10 (human-IgG)
Usable temperature 4-60°C
Usable pH range 2-12
Column material PEEK
POINTS
  • Newly developed hydrophilic polymer beads with low nonspecific adsorption
  • Effective surface structure designed to interact with biomolecules
  • High theoretical plate number and symmetrical peak shape by optimal packing technology
  • QA (quaternary ammonium) and SP (sulfopropyl) ion exchangers
  • Ideal for analysis and laboratory-scale purification on porous type with high binding capacity and high recovery of biomolecules
  • Ultra-fast analysis and high-resolution analysis on non-porous type

APPLICATION DATA

Features

Monoclonal antibody (MAb) analysis on non-porous type cation exchange columns

The separation of MAb is compared on BioPro IEX SF and competitor's column under the same gradient conditions at pH 5.6.
BioPro IEX SF column provides higher resolution of MAb in a shorter analysis time than the competitor's column.

Eluent A) 20 mM MES-NaOH (pH 5.6)
B) 20 mM MES-NaOH (pH 5.6) containing 0.2 M NaCl
Initial gradient conc. 35%B (70 mM NaCl)
Gradient slope 0.25%B/min (0.5 mM NaCl)
Flow rate 180 cm/hr
(0.5 mL/min for 100 X 4.6 mmI.D.,
0.378 mL/min for 250 X 4.0 mmI.D.)
Temperature 30°C
Detection UV at 280 nm
Injection 10 µL
Sample Humanized monoclonal IgG1 (1 mg/mL)

Excellent batch-to-batch reproducibility

BioPro IEX SF column exhibits excellent batch-to-batch reproducibility for MAb analysis, including the resolution of peaks for small charge variants. All the gel batches are inspected by various quality control tests, including HPLC analysis of MAb, and must pass rigorous criteria before release. BioPro IEX Columns are the best choice for the quality control of MAb and other biopharmaceuticals.

Column BioPro IEX SF 5 µm, 100 X 4.6 mmI.D.
Eluent A) 20 mM NaH2PO4-Na2HPO4  (pH 6.5)
B) 20 mM NaH2PO4-Na2HPO4  (pH 6.5) containing 0.2 M NaCl
0-50%B (0.5-30 min)
Flow rate 0.5 mL/min (180 cm/hr)
Temperature 25°C
Detection UV at 215 nm
Injection 10 µL
Sample Humanized monoclonal IgG1

30 mm-length column for high-throughput analysis with low operating pressure

The separation performance of non-porous and porous columns for proteins is compared. BioPro IEX Colomn non-porous types are effective for high-throughput analysis at a high flow rate because of its high mechanical strength and high column efficiency.

  1. Ribonuclease A
  2. Cytochrome c
  3. Lysozyme
Eluent A) 20 mM KH2PO4-K2HPO4 (pH 6.8)
B) 20 mM KH2PO4-K2HPO4 (pH 6.8)
    containing 0.5 M NaCl
0-100%B (0-4 min) for BioPro IEX SF
0-100%B (0-60 min) for BioPro IEX SP
Temperature 25°C
Detection UV at 220 nm
Injection 20 µL

High binding capacity and high recovery of biomolecules

BioPro IEX QA gives the superior DBC and recovery compared with conventional porous polymer anion exchange columns. The surface structure of porous type BioPro IEX, which is designed for maximum interaction with proteins, provides high binding capacity, and the hydrophilic property of polymer beads significantly reduces nonspecific adsorption of proteins


Comparisons of dynamic binding capacity (DBC) and recovery for BSA

  Dynamic binding capacity
(mg/mL-resin, 10% breakthrough)
Eluted amount
(mg/mL-resin)
Recovery*
(%)
BioPro IEX QA 126 120 95
Brand T (porous Q type) 73 58 79
Brand G (porous Q type) 100 35 35

*Recovery: (Eluted amount/DBC) X 100

Breakthrough curves

Column BioPro IEX QA, 50 X 4.6 mmI.D.
Brand T (porous Q Type) 50 X 4.6 mmI.D.
Brand G (porous Q Type) 50 X 5.0 mmI.D.
Linear velocity 180 cm/hr
Equilibration buffer 20 mM Tris-HCl (pH 8.6)
Elution buffer 20 mM Tris-HCl (pH 8.6) containing 1.0 M NaCl
Detection UV at 280 nm
Sample 1 mg/mL Bovine serum albumin (BSA) in
equilibration buffer

High loadability

BioPro IEX QA shows excellent resolution and peak shapes even when the loading amount increases. The porous type BioPro IEX columns are suitable for laboratory-scale purification of proteins.

Comparison of the effect of sample load on BioPro IEX QA and commercial Q type column

  1. Ovalbumin
  2. Trypsin inhibitor
Eluent A) 20 mM Tris-HCl (pH 8.1)
B) 20 mM Tris-HCl (pH 8.1)
    containing 0.5 M NaCl
10-80%B (0-30 min)
Flow rate 0.5 mL/min (180 cm/hr for 4.6 mmI.D., 150 cm/hr for 5.0 mmI.D.)
Temperature 25°C
Detection UV at 280 nm
Injection 100 µL