期刊
ANALYTICAL CHEMISTRY
卷 78, 期 17, 页码 6265-6269出版社
AMER CHEMICAL SOC
DOI: 10.1021/ac060251b
关键词
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资金
- NIGMS NIH HHS [GM 59240] Funding Source: Medline
A new algorithm, sequence-specific retention calculator, was developed to predict retention time of tryptic peptides during RP HPLC fractionation on C18, 300-angstrom pore size columns. Correlations of up to similar to 0.98 R-2 value were obtained for a test library of (similar to) 2000 peptides and similar to 0.95-0.97 for a variety of real samples. The algorithm was applied in conjunction with an exclusion protocol based on mass ( 15 ppm tolerance) and retention time (2-min tolerance for 0.66% acetonitrile/min gradient), MART criteria to significantly reduce the instrument time required for complete MS/MS analysis of a digest separated by RP HPLC. This was confirmed by reanalyzing the set of HPLC-MALDI MS/MS data with no loss in protein identifications, despite the number of virtually executed MS/MS analyses being decreased by 57%.
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