Enhanced Detection of Low Abundance Human Plasma Proteins Using a Tandem IgY12-SuperMix Immunoaffinity Separation Strategy

• DOI: 10.1074/mcp.m800008-mcp200
• Publication Date: 2008-07-17T00:14:54Z
• Authors: Wei-Jun Qian; David T. Kaleta; Brianne O. Petritis; Hongliang Jiang; Tao Liu; Xu Zhang; Heather M. Mottaz; Susan M. Varnum; David G. Camp; Lei Huang; Xiangming Fang; Wei-Wei Zhang; Richard D. Smith
• Keywords: Tandem

The enormous dynamic range of human bodily fluid proteomes poses a significant challenge for current MS-based proteomics technologies as it makes especially difficult to detect low abundance proteins in biofluids such blood plasma, which is an essential aspect successful biomarker discovery efforts. Here we present novel tandem IgY12-SuperMix immunoaffinity separation system enhanced detection plasma. separates approximately 60 abundant from the allowing enrichment plasma SuperMix flow-through fraction. High reproducibility separations was observed terms both sample processing recovery and LC-MS/MS identification results based on spectral count data. ability quantitatively measure differential protein abundances following application demonstrated by spiking six non-human standard at three different levels into A side-by-side comparison between IgY12 samples analyzed one- two-dimensional revealed 60-80% increase proteome coverage result separations, suggesting significantly proteins. total 695 were confidently identified single analysis (with minimum two peptides per protein) coupling strategy with LC-MS/MS, including 42 reported normal concentrations 100 pg/ml ng/ml. selected proteins, macrophage colony-stimulating factor 1 matrix metalloproteinase-8, independently validated ELISA 202 12.4 ng/ml, respectively. Evaluation binding efficiency that 45 medium efficiently captured column >90% retention. Taken together, these illustrate potential broad utilities this applications involving where effectively addressing specimen imperative.