Evaluation of multiprotein immunoaffinity subtraction for plasma proteomics and candidate biomarker discovery using mass spectrometry.
Strategies for removal of high abundance proteins have been increasingly utilized in proteomic studies of serum/plasma and other body fluids to enhance the detection of low abundance proteins and achieve broader proteome coverage; however, both the reproducibility and specificity of the high abundance protein depletion process still represent common concerns. Here we report a detailed evaluation of immunoaffinity subtraction performed applying the ProteomeLab IgY-12 system that is commonly used in human serum/plasma proteome characterization in combination with high resolution LC-MS/MS. Plasma samples were repeatedly processed using this approach, and the resulting flow-through fractions and bound fractions were individually analyzed for comparison. The removal of target proteins by the immunoaffinity subtraction system and the overall process was highly reproducible. Non-target proteins, including one spiked protein standard (rabbit glyceraldehyde-3-phosphate dehydrogenase), were also observed to bind to the column at different levels but also in a reproducible manner. The results suggest that multiprotein immunoaffinity subtraction systems can be readily integrated into quantitative strategies to enhance detection of low abundance proteins in biomarker discovery studies.
Liu
Tao
T
Biological Sciences Division and Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington 99352, USA.
Qian
Wei-Jun
WJ
Mottaz
Heather M
HM
Gritsenko
Marina A
MA
Norbeck
Angela D
AD
Moore
Ronald J
RJ
Purvine
Samuel O
SO
Camp
David G
DG
Smith
Richard D
RD
D015415:Biomarkers / Q000032:analysis; D001798:Blood Proteins / Q000032:analysis* / Q000276:immunology; D002846:Chromatography, Affinity / Q000295:instrumentation*; D006801:Humans; D007136:Immunoglobulins / Q000276:immunology; D020543:Proteome / Q000032:analysis* / Q000276:immunology; D040901:Proteomics / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D015415:Biomarkers; D001798:Blood Proteins; C016512:IgY; D007136:Immunoglobulins; D020543:Proteome