Top Down and Accurate Mass Analysis of Complex Protein Mixtures with Improved Chromatographic Performance

摘要

1) Chromatographic separation of more than 15 ribosomal proteins was achieved using 5 um polymeric reversed phase material (Polymer Laboratories, Inc.) packed into a 75um fused silica column. 2) Intact protein masses were determined from SIM scans of the precursors with 100,000 resolution using the Orbitrap MS. 3) Sequence tags were generated from the deconvoluted CID fragments of multiply charged precursors. 4) A correct protein identification was achieved using the sequence tag generated from one of the deconvoluted CID spectra. 5) Greater than 99percent sequence coverage was achieved when based on peptide fragment masses.