HPLC Analysis of the Monoclonal Antibody (mAb) Erbitux (Cetuximab) on BIOshell™ A400 Protein C4
Conditions | |
|---|---|
| Column | BIOshell™ A400 Protein C4, 10 cm X 2.1 mm I.D., 3.4 µm particles (66825-U) |
| Column temp. | 90 °C |
| Mobile phase | (A) 80:20, 0.1% TFA in water: 0.1% TFA in acetonitrile; (B) 20:80, 0.1% TFA in water: 0.1% TFA in acetonitrile |
| Gradient | 20 to 35%B in 10 min (or 5 min) |
| Flow rate | 0.2 mL/min |
| Pressure | 470 psi (or 940 psi) |
| Sample | 1 g/L mAb in 20 mM Tricine/KOH, pH 7 |
| Injection | 0.4 µL |
| Detector | UV, 215 nm |
Description | |
|---|---|
| Analysis Note | Antibodies can exhibit considerable hydrophobicity, and so efficient reversed-phase elution of this intact mAb requires the use of high temperature. The top chromatogram illustrates that BIOshell™ Fused-Core® columns can be operated at high flow rate while maintaining good peak shape at a reasonable back pressure. |
| Categories | Analytical Chromatography, Proteins |
| Featured Industry | Life Science and Biopharma |
| Legal Information | BIOshell is a trademark of Sigma-Aldrich Co. LLC, Fused-Core is a registered trademark of Advanced Materials Technology, Inc. |
| Suitability | Application for HPLC |
Materials
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