UHPLC Analysis of SigmaMAb Universal Antibody Standard on BIOshell IgG 1000 Å C4
CONDITIONS
column
BIOshell IgG 1000 Å C4; 10 cm x 2.1 mm I.D., 2.7 μm particles (63288-U)
mobile phase
[A] 70:30 0.1% TFA (water): 0.1% TFA (acetonitrile); [B] 50:50 0.1% TFA (water): 0.1% TFA (acetonitrile)
gradient
0% B to 50% B in 50 min; 50% B to 100% B in 10 min
flow rate
0.2 mL/min
pressure
1325 psi (91.4 bar)
column temp.
75 °C
detector
UV, 215 nm
injection
1.0 μL
sample
SigmaMAb (p/n MSQC4), 1 g/L, 0.05% TFA (water)
Description
Analysis Note
The BIOshell IgG column is uniquely optimized for the resolution of analytes with high molecular weights like proteins and antibodies. The SigmaMAb Universal Antibody Standard is a recombinant human monoclonal antibody with a molecular mass of ~150 kDa expressed in Chinese Hampster Ovary (CHO) cells. This application demonstrates the ability of the BIOshell IgG column to resolve impurities and other species within the main analyte peak. Superior resolution was attained on the BIOshell IgG column, along with better recovery of the antibody, over a conventional, 1.7 μm UHPLC column packed with fully porous particles (FPPs).
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