UNIT 24.1 Human Serum and Plasma Proteomics

Abstract

This unit focuses on several related methods for profiling and identifying proteins in human serum or plasma. While the methods are specifically described for human serum and plasma, with the possible exception of the major protein depletion steps, they can be readily adapted to serum and plasma from other species as well as to other biological fluids. A very powerful multidimensional protein-profiling method is described that utilizes major protein depletion coupled to two other protein separations, MicroSol IEF fractionation and SDS-PAGE, followed by LC-MS/MS. Other protein-profiling strategies include the depletion of abundant proteins followed by analysis either via 2-D gels or using offline Multidimensional Protein Identification Technology (MudPIT) or LC/LC-MS/MS. Additional protein profiling methods and strategies are also discussed.