Evaluation of Reversed-Phase Columns for Protein Separation Using a Single LC/MS - LC/UV Methodology

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Kurt Yardley, Paul Rainville, Jeff Mazzeo, Uwe Neue, Reb Russell II
HPLC 2002; Montreal; 5-6 June 2002
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HPLC separations on reversed phase columns are an important part of characterizing complex protein samples from basic research and pharmaceuticals. Recent advances in mass spectrometry have enabled rapid characterization of protein and peptide samples separated by HPLC, greatly facilitating the identification of target proteins and contaminants. Thus, there is a need for good separation techniques using MS compatible methodology for the separation of proteins by HPLC. We present here the evaluation of peak capacity and recovery in protein separations along with LC/MS. Our findings resulted in the determination that both Delta-Pak and Symmetry300 columns are preferred for performing LC/MS protein separations.

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