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NEW SCX PEPTIDE ELUTION SCORE FOR PH/SALT-GRADIENT SCX CHROMATOGRAPHY IN 2D-NANO-LC/MSMS ANALYSIS OF PROTEIN DIGESTS
Citation:
WINNIK, W. M. NEW SCX PEPTIDE ELUTION SCORE FOR PH/SALT-GRADIENT SCX CHROMATOGRAPHY IN 2D-NANO-LC/MSMS ANALYSIS OF PROTEIN DIGESTS. Presented at American Society for Mass Spectrometry, San Antonio, TX, June 05 - 09, 2005.
Description:
A new automated 2D-(SCX/RP)-nano-LC/MSMS method was developed. Separation of the peptides in the first LC dimension was the main focus of this work, and it was optimized using human serum albumin (HSA) and human lung cell lysate tryptic digests. Samples were reduced and alkylated with different cysteine alkylating reagents to provide peptides possessing identical amino acid sequences but different cysteine tags. In particular, a quaternary amine tag (APTA) was useful for the evaluation of peptides elution order under SCX conditions. The optimum SCX peptide elution conditions involved a simultaneous pH - gradient with ammonium acetate salt - gradient elution. A new SCX Peptide Elution Score was developed that correlates with the elution pH.