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HPLC Analysis of SILu Lite Antibody Standard on BIOshell™ A400 Protein C4 Versus Competitor Column

HPLC Analysis of SILu Lite Antibody Standard on BIOshell™ A400 Protein C4 Versus Competitor Column

Conditions

column BIOshell A400 Protein C4, 10 cm x 2.1 mm I.D., 3.4 µm particles (66825-U)
column Competitor Column, 10 cm x 2.1 mm I.D., 1.8 µm particles
column temp. 75 C
mobile phase [A] 70:30 water (0.1% trifluoroacetic acid): acetonitrile (0.1% trifluoroacetic acid); [B] 50:50 water (0.1% trifluoroacetic acid): acetonitrile (0.1% trifluoroacetic acid)
gradient 0 to 100% B in 12 min; held at 100% B for 2 min
flow rate 0.3 ml/min
pressure 860 psi (59 bar)
sample SILu Lite Antibody Standard, 1 g/L, 0.05% trifluoroacetic acid
injection 1 µl
detector UV, 215 nm

Description

Analysis Note The BIOshell A400 Protein C4 column is designed for efficient separations of large proteins and peptides that may be too well-retained on a traditional C18 phase. An antibody standard was assayed on the BIOshell A400 C4 column and on a comparable competitor column with a sub-2 μm particle size. The assay was performed on the same day using the same HPLC sample. The top chromatogram shows the antibody standard that was run on the BIOshell column while the bottom chromatogram shows the antibody standard that was run on the competitor column. The BIOshell A400 C4 column provided better resolution of minor components than the competitor column and provided a narrower peak width for the analyte peak. The 3.4 μm particle size of the BIOshell A400 C4 column had a much lower operating pressure than the sub-2 μm competitor column (860 psi versus 2460 psi), thus allowing this separation to take place under non-UHPLC conditions.
Categories Analytical Chromatography, Proteins, Peptides
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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