Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

JoVE 2812 7/31/2011

Lei Zhao*¹, Jeffrey R. Whiteaker*¹, Matthew E. Pope², Eric Kuhn³, Angela Jackson⁴, N. Leigh Anderson⁵, Terry W. Pearson², Steven A. Carr³, Amanda G. Paulovich¹

¹Clinical Research Division, Fred Hutchinson Cancer Research Center - FHCRC, ²Department of Biochemistry and Microbiology, University of Victoria, ³Broad Institute of MIT and Harvard, ⁴Genome BC Proteomics Centre, University of Victoria, ⁵Plasma Proteome Institute

Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA) couples affinity enrichment of peptides with stable isotope dilution mass spectrometry (MRM-MS) to provide quantitative measurement of peptides as surrogates for their respective proteins. Here we describe the protocol using magnetic particles in a partially automated format.