Isolation of Major Histocompatibility Complex (MHC)-Associated Peptides by Immunoaffinity Purification
MHC molecules on the surface of cells are responsible for the presentation of antigenic peptides to CD8+ and CD4+ T cells. Downstream analysis of these peptides can offer insight into various disease processes with an immune component, as in autoimmune diseases and cancer. A critical step in studying MHC-associated peptides is their isolation from MHC molecules on the surface of cells. In this chapter, we detail anti-MHC antibody conjugation to beads, immunoaffinity purification of the MHC molecules, and peptide elution. This method can be used for analysis of unmodified and/or posttranslationally modified peptides by mass spectrometry.
Key wordsMHC-associated peptide isolation Immunoaffinity purification Peptide elution
The authors would like to acknowledge Mark Cobbold (MGH) and Donald F. Hunt (U Virginia) for their technical and financial support. S.A.M. is currently funded by an NIH F32 postdoctoral fellowship.
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