HPLC Analysis of Oxidized SILu Lite SigmaMAb Universal Antibody Standard on BIOshell A400 Protein C4

HPLC Analysis of Oxidized SILu Lite SigmaMAb Universal Antibody Standard on BIOshell A400 Protein C4

Conditions

instrument Thermo Ultimate BioRS 3000
column BIOshell A400 Protein C4, 10 cm x 2.1 mm I.D., 3.4 µm
mobile phase [A] 70:30, (0.1% TFA in water):(0.1% TFA in acetonitrile; [B] 50:50,  (0.1% TFA in water):(0.1% TFA in acetonitrile)
gradient 0 to 50% B in 50 min; 50 to 100% B in 10 min
flow rate  0.2 mL/min
pressure 580 psi
column temp. 75 °C
detector UV, 215 nm
injection 1.0 µL

Description

Featured Industry Life Science and Biopharma 
Legal Information BIOshell is a trademark of Sigma-Aldrich Co. LLC
SILu is a trademark of Sigma-Aldrich Co. LLC
Analysis Note This application describes conditions for the separation and detection of oxidized variants of a universal antibody standard by reversed-phase chromatography.  Several amino acids, such as tryptophan, methionine, and cysteine, are susceptible to oxidation. As components of proteins, when these amino acids are oxidized, various deleterious effects to the protein may ensue, that affect structure and/or function.  Biotherapeutics are susceptible to protein oxidation; therefore, monitoring a product mixture for oxidation variants is paramount.  The top panel of the figure shows the native SigmaMAb antibody standard while the bottom panel shows the SigmaMAb antibody standard after oxidative stress with hydrogen peroxide.
Application No. ASC120118A

Materials

     
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