HPLC Analysis of the Monoclonal Antibody (mAb) Erbitux (Cetuximab) on BIOshell™ A400 Protein C4

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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