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HPLC of monoclonal antibody (mAb) SigmaMab on BIOshell™ A400 Protein C4

HPLC of monoclonal antibody (mAb) SigmaMab 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 (or 0.4 ml/min)
pressure 470 psi (or 940 psi) (32 bar or 65 bar)
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 high temperature. With the BIOshell Fused-Core particle column, high flow rates can be applied while retaining good separation, while still operating at reasonable column backpressure.
Featured Industry Life Science and Biopharma
Legal Information BIOshell is a trademark of Sigma-Aldrich Co. LLC
suitability application for HPLC

Materials

     
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