Journal
BIOTECHNIQUES
Volume 62, Issue 3, Pages 123-130Publisher
FUTURE SCI LTD
DOI: 10.2144/000114525
Keywords
peptide; major histocompatibility complex (MHC); pMHC; dextramer; T-cell receptor (TCR); immunology; cancer immunotherapy; ligand discovery; tumor antigen discovery; peptide-specific MHC clustering; multimer; tetramer; biotinylated dextran
Funding
- National Institutes of Health [5P01CA132681-5]
- Prostate Cancer Foundation Challenge Award [15CHAL02]
- Jane Coffin Childs Postdoctoral Fellowship
- NIH grant [R35 CA197633]
- Ressler Family Foundation
Ask authors/readers for more resources
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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available