Elimination of affinity reagent interference for the mass spectrometric detection of low-abundance proteins following immunoprecipitation.
The presence of affinity reagents such as immunoglobulin in preparations for sensitive mass spectrometry analyses can preclude the identification of low-abundance proteins of interest. We report a method whereby antisera are purified and biotinylated prior to use in immunoprecipitation that allows for its efficient removal from proteomic samples via streptavidin capture. This method can similarly be extended to other affinity reagents such as recombinant fusion proteins for enhanced identification of interacting proteins.
Martin
Angela M
AM
Department of Microbiology, Immunology, and Molecular Genetics and Department of Chemistry, University of Kentucky, Lexington, Kentucky 40536, USA.
Liu
Ting
T
Lynn
Bert C
BC
Sinai
Anthony P
AP
D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport; D007136:Immunoglobulins / Q000378:metabolism; D047468:Immunoprecipitation; D007202:Indicators and Reagents; D008565:Membrane Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011485:Protein Binding / Q000502:physiology; D053719:Tandem Mass Spectrometry; D014122:Toxoplasma / Q000378:metabolism; D014617:Vacuoles / Q000737:chemistry / Q000378:metabolism
D007136:Immunoglobulins; D007202:Indicators and Reagents; D008565:Membrane Proteins