HPLC Analysis of the Monoclonal Antibody (mAb) Erbitux (Cetuximab) on BIOshell™ A400 Protein C4
Materials
related product
used together
BIOshell™ A400 Protein C4, 3.4 μm Guard Cartridge
3.4 μm particle size, L × I.D. 0.5 cm × 2.1 mm, pkg of 3 eaCONDITIONS
column
BIOshell™ A400 Protein C4, 10 cm X 2.1 mm I.D., 3.4 μm particles (66825-U)
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)
column temp.
90 °C
detector
UV, 215 nm
injection
0.4 μL
sample
1 g/L mAb in 20 mM Tricine/KOH, pH 7
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.
Legal Information
BIOshell is a trademark of Sigma-Aldrich Co. LLC, Fused-Core is a registered trademark of Advanced Materials Technology, Inc.