Use of Peptide Retention Time Prediction for Protein Identification by off-line Reversed-Phase HPLC−MALDI MS/MS
| Autor: | John P. Cortens, Werner Ens, John A. Wilkins, Stephen M. Ying, Victor Spicer, Dhimankrishna Ghosh, Kenneth G. Standing, Oleg V. Krokhin, Ronald C. Beavis |
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| Rok vydání: | 2006 |
| Předmět: |
Ions
chemistry.chemical_classification Pore size Time Factors Chromatography Chemistry Analytical chemistry Proteins Peptide Fractionation Reversed-phase chromatography Mass spectrometry Online Systems High-performance liquid chromatography Analytical Chemistry Matrix-assisted laser desorption/ionization Tandem Mass Spectrometry Cell Line Tumor Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Humans Databases Protein Peptides Retention time Chromatography High Pressure Liquid |
| Zdroj: | Analytical Chemistry. 78:6265-6269 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/ac060251b |
| Popis: | A new algorithm, sequence-specific retention calculator, was developed to predict retention time of tryptic peptides during RP HPLC fractionation on C18, 300-A pore size columns. Correlations of up to approximately 0.98 R2 value were obtained for a test library of approximately 2000 peptides and approximately 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%. |
| Databáze: | OpenAIRE |
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