Biochromatography
YMC-BioPro
YMC-BioPro series ion exchange columns are for separation of proteins, peptides, and nucleic acids. YMC-BioPro IEC columns are available in QA and SP chemistries and are based on 5 micron porous (QA and SP columns) and non-porous (QA-F and SP-F 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
| YMC-BioPro QA | YMC-BioPro SP | YMC-BioPro QA-F | YMC-BioPro SP-F | |
|---|---|---|---|---|
| Matrix | hydrophilic porous polymer beads | hydrophilic non-porous polymer beads | ||
| Particle size | 5µm | |||
| Charged group | -CH2N+(CH3)3 | -CH2CH2CH2SO3- | -CH2N+(CH3)3 | -CH2CH2CH2SO3- |
| Counter ion | Cl- | Na+ | Cl- | Na+ |
| Ion-exchange capacity | 0.075-0.100 meq/mL-resin |
0.070-0.095 meq/mL-resin |
0.075-0.110 meq/mL-resin |
0.230-0.290 meq/mL-resin |
| Dynamic binding capacity | >110 mg-BSA/ mL-resin |
>70 mg-human-IgG/ mL-resin |
>12 mg-BSA/ mL-resin |
>10 mg-human-IgG/ mL-resin |
| Available pH range | 2-12 | |||
| Column material | PEEK | |||
POINT
- 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
Excellent resolution
- Ribonuclease A (0.5 mg/mL)
- Cytochrome c (0.5 mg/mL)
- Lysozyme (0.5 mg/mL)
| 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 (0-60 min) |
|---|---|---|
| Flow rate | : | 0.5 mL/min for 4.6 mmI.D. 0.59 mL/min for 5.0 mmI.D. |
| Temperature | : | 25℃ |
| Detection | : | UV at 220 nm |
| Injection | : | 20µL for 4.6 mmI.D. 23.6µL for 5.0 mmI.D. |
The separation patterns of standard protein mixtures are compared among YMC-BioPro SP and two conventional porous-polymer cation-exchange columns.
Most of impurities are resolved from the main peaks of proteins on YMC-BioPro SP with superior peak shapes.
High binding capacity and high recovery of biomolecules
Comparisons in dynamic binding capacity (DBC) and recovery for BSA
| Dynamic binding capacity (mg/mL-resin, 10% breakthrough) |
Eluted amount (mg/mL-resin) |
Recovery* (%) |
|
|---|---|---|---|
| YMC-BioPro QA | 126 | 120 | 95 |
| Brand T (porous Q type) | 73 | 58 | 79 |
| Brand G (porous Q type) | 100 | 35 | 35 |
*Recovery: (Eluted amount/Dynamic binding capacity) X 100
Breakthrough curves
| Column | : | YMC-BioPro 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 | : | 3.0 cm/min |
| Sample | : | 1 mg/mL Bovine serum albumin (BSA) in equilibration buffer |
| 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 |
Compared with conventional porous-polymer anion-exchange columns, YMC-BioPro QA gives a superior DBC. Furthermore, the recovery is greater than that of conventional columns. The hydrophilic properties of the matrix polymer remarkably reduce nonspecific adsorption of proteins on YMC-BioPro columns.
High loadability
Excellent peak shapes even at high sample load
- Ovalbumin
- 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 |
| Temperature | : | 25℃ |
| Detection | : | UV at 280 nm |
| Injection | : | 100µL |
YMC-BioPro QA shows the excellent peak shapes even when the loading amount increases. By contrast, the column of Brand G cannot achieve acceptable peak shapes and resolution even in small amount of injection.
30 mm-length column for high-throughput analysis with low operating pressure
- Ribonuclease A
- Cytochrome c
- 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 YMC-BioPro SP-F 0-100%B (0-60 min) for YMC-BioPro SP |
|---|---|---|
| Temperature | : | 25℃ |
| Detection | : | UV at 220 nm |
| Injection | : | 20µL |
The separation performance of proteins is compared between non-porous column and porous column. YMC-BioPro non-porous types are effective for high-throughput analysis with high flow rate because of high mechanical strength and high column efficiency.
100 mm-length column for high-resolution analysis
| Eluent | : | A) 20 mM Tris-HCl (pH 8.1) B) 20 mM Tris-HCl (pH 8.1) containing 0.5 M NaCl 10-25%B (0-60 min) |
|---|---|---|
| Flow rate | : | 1.0 mL/min |
| Temperature | : | 25℃ |
| Detection | : | UV at 220 nm |
| Injection | : | 14µL (0.1 mg/mL) |
| Sample | : | Mouse monoclonal IgG1 anti-human IgG4 (Purified by DEAE chromatography, containing NaN3) |
Two different lots of commercially available MAb, purified by DEAE chromatography, are separated with 100 mm-length of YMC-BioPro QA-F column.
The MAb is resolved into several peaks and the lot-to-lot variability is observed.
The 100 mm-length columns of YMC-BioPro QA-F and SP-F have high efficiency. They are ideal for characterization or QC assessment of closely related proteins such as Mab isoforms.