Protocol for cell surface biotinylation of magnetic labeled and captured human peripheral blood mononuclear cells

STAR Protoc. 2022 Dec 16;3(4):101863. doi: 10.1016/j.xpro.2022.101863. Epub 2022 Nov 21.


Analysis of the surfaceome of a blood cell subset requires cell sorting, followed by surface protein enrichment. Here, we present a protocol combining magnetically activated cell sorting (MACS) and surface biotinylation of the target cell subset from human peripheral blood mononuclear cells (PBMCs). We describe the steps for isolating target cells and their in-column surface biotinylation, followed by isolation and mass spectrometry analysis of biotinylated proteins. The protocol enables in-column surface biotinylation of specific cell subsets with minimal membrane disruption.

Keywords: Cell Biology; Cell Isolation; Mass Spectrometry; Protein Biochemistry; Proteomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotinylation
  • Cell Membrane / metabolism
  • Humans
  • Leukocytes, Mononuclear* / chemistry
  • Magnetic Phenomena
  • Membrane Proteins* / metabolism


  • Membrane Proteins