简介：

Here we describe a method for extensive protein identification in complex samples that involves iterative nanoflow LC-MS/MS analysis, exclusion list generation, and iterative database searching with Thermo Scientific Proteome Discoverer software, leading to the identification of numerous low-copy-number proteins in the seminal plasma proteome.

仪器：

Thermo Scientific LTQ Orbitrap XL

总结：

The combination of protein enrichment and iterative injections with exclusion of previously identified peptides resulted in the identification of many more peptides and proteins in the seminal plasma sample. Re-injection of the same sample allowed the identification of about 80% new peptides in each run compared to about 25% without exclusion lists. New proteins were identified, “one hit wonder” proteins could be confirmed by additional peptides, and overall the protein coverage was increased, demonstrating the benefit of this strategy for deep proteomic characterization of complex biological samples such as seminal plasma.