P36-T Characterization of the Analytical Performance of an FT-MS-Based Label-Free Platform for Biomarker Discovery in Human Plasma: Standards, Precision, Accuracy, and Directed Feature Investigation Using SIEVE

In this report we present a detailed methodological and instrumental description of an FT-MS-based, label-free platform for biomarker discovery in human plasma. We will discuss the relevant analytical figures of merit for each of the main steps of this workflow, namely immunoaffinity depletion of the 12 most abundant plasma proteins; high-precision and high-resolution LC-MS/MS analysis using a unique split-flow design; robust differential MS signal analysis using a robust high-throughput computational assembly that employs new chromatographic alignment and time-course statistics-based differential expression algorithms (SIEVE), followed by directed feature (frame) identification using Sequest. The application of this analytical platform to a relevant clinical model will be discussed, as well as its extension to targeted feature analysis by complementary MS strategies (ECD, IRMPD), and potential new applications such as metabolomics.