UHPLC Analysis of Human IgM on BIOshell(TM) IgG 1000 Ã… C4

UHPLC Analysis of Human IgM on BIOshell™ IgG 1000 Ã… C4

Conditions

column BIOshell IgG 1000 Ã… C4, 10 cm x 2.1 mm I.D., 2.7 μm particles (63288-U)
column temp. 75 °C
mobile phase [A] 0.1% TFA (water); [B] 0.1% TFA (acetonitrile)
gradient 30% B to 60% B in 10 min; 60% B to 100% B in 10 min
flow rate 0.2 mL/min
pressure 1425 psi (98.3 bar)
sample IgM, 100 μg/mL, 0.1% TFA (water)
injection 1 μL
detector UV, 215 nm

Description

Analysis Note This application demonstrates the advantage of using a larger pore size to analyze large analytes, in particular, biomacromolecules. IgM is a pentamer with a molecular weight of ~970 kDa; therefore, a much larger pore size is required to attain adequate peak shape and recovery. This is shown in the above chromatograms: the top trace, which employs the 1000 Ã… pore size BIOshell IgG column displays narrower peak shape and better recovery than a fully porous, 300 Ã… competitor column.
Featured Industry Life Science and Biopharma
Legal Information BIOshell is a trademark of Sigma-Aldrich Co. LLC
suitability application for UHPLC

Materials

     
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