Preparation of peptide-MHC and T-cell receptor dextramers by biotinylated dextran doping

Biotechniques. 2017 Mar 1;62(3):123-130. doi: 10.2144/000114525.


Peptide-major histocompatibility complex (pMHC) multimers enable the detection, characterization, and isolation of antigen-specific T-cell subsets at the single-cell level via flow cytometry and fluorescence microscopy. These labeling reagents exploit a multivalent scaffold to increase the avidity of individually weak T-cell receptor (TCR)-pMHC interactions. Dextramers are an improvement over the original streptavidin-based tetramer technology because they are more multivalent, improving sensitivity for rare, low-avidity T cells, including self/tumor-reactive clones. However, commercial pMHC dextramers are expensive, and in-house production is very involved for a typical biology research laboratory. Here, we present a simple, inexpensive protocol for preparing pMHC dextramers by doping in biotinylated dextran during conventional tetramer preparation. We use these pMHC dextramers to identify patient-derived, tumor-reactive T cells. We apply the same dextran doping technique to prepare TCR dextramers and use these novel reagents to yield new insight into MHC I-mediated antigen presentation.

Keywords: T-cell receptor (TCR); biotinylated dextran; cancer immunotherapy; dextramer; immunology; ligand discovery; major histocompatibility complex (MHC); multimer; pMHC; peptide; peptide-specific MHC clustering; tetramer; tumor antigen discovery.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotechnology / methods*
  • Biotin
  • Dextrans / chemistry
  • Dextrans / metabolism*
  • Flow Cytometry
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism
  • HEK293 Cells
  • Histocompatibility Antigens / chemistry
  • Histocompatibility Antigens / genetics
  • Histocompatibility Antigens / metabolism*
  • Humans
  • Jurkat Cells
  • K562 Cells
  • Peptides / chemistry
  • Peptides / genetics
  • Peptides / metabolism*
  • Receptors, Antigen, T-Cell / chemistry
  • Receptors, Antigen, T-Cell / genetics
  • Receptors, Antigen, T-Cell / metabolism*
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism*
  • Streptavidin
  • Transduction, Genetic


  • Dextrans
  • Fluorescent Dyes
  • Histocompatibility Antigens
  • Peptides
  • Receptors, Antigen, T-Cell
  • Recombinant Fusion Proteins
  • Biotin
  • Streptavidin