MHC Class I Tetramers

Location: Thomas Building, DE-550
Contact phone: (206) 667-6455
Contact fax: (206) 667-6845
Contact e-mail: jcao@fredhutch.org

pMHC tetramers are multimeric peptide-MHC complexes that serve as reagents for analyzing antigen-specific T cells. The multimeric nature of tetramers increases the affinity between pMHC and TCR enabling their usage as labeling reagents for TCR. Labeling of antigen-specific T cells by tetramers makes possible the characterization of the phenotype and functionality of these cell populations.

Potential use of pMHC Tetramers

  • Enumeration of antigen-specific T cells
  • Characterization of antigen-specific T cells
  • Isolation of antigen-specific T cells
  • In situ staining of antigen-specific T cells

In the production of tetramers, the desired MHC heavy chain protein and epitopic peptide are refolded into MHC complex with 2-microglobulin in vitro. The successful refolded monomers are purified by gel-filtration and then biotinylated. Tetramerization is achieved by binding to streptavidin, a fluorescent dye often added by using fluorochrome-conjugated streptavidin.

The Immune Monitoring resource can produce pMHC monomers and tetramers with defined MHC alleles and epitopic peptides. We also have available many monomers and tetramers available that have been previously produced. See our product information and list or contact the facility for more details.

Note: Individual tetramers can differ significantly in refolding, stability and staining of T cells. The differences are determined largely by the HLA alleles and the binding affinity of the specific peptide to the HLA molecule. We recommend the testing of each new batch of tetramers with positive and negative control cells and by titration of the tetramer reagent. Tetramers should be stored at 4°C and retested after long-term storage. Please visit the NIH Tetramer Core Facility's website for more information.

Clinical Grade Tetramers

We produce clinical grade pMHC Class-I tetramers. These custom-made, sterile and highly purified products are used to sort antigen-specific CD8+ T cells, which are cultured and then used for immune therapy trials.  The production of these tetramers is approved by the FDA to use in certain IND protocols at Fred Hutch.

Requesting Service

For any of the above assays please use iLab to request the service.