MHC multimer staining for detection of antigen-specific T cells

We believe the most powerful strategy for mapping antigen-specific T cells is staining with MHC multimers.

Common MHC multimer staining

Our specialty within detection of antigen-specific T cell responses and the most exact method for enumerating and profiling these is the MHC multimer staining for flow cytometry. In addition to staining with the MHC multimers we co-stain for T cell lineage markers and often further characterize the antigen-specific T cells in regard to e.g. their maturation phenotype and further markers. For an example see our slide on Deep Profiling of Antigen-specific T cells.

Upon treatment with immunotherapies in human or murine cohorts, the enumeration and activation of certain antigen-specific T cells is important to evaluate the treatment-induced immune response by comparing pre- and post-treatment samples.
Multiple different antigen-specific T cells populations can be detected using MHC multimer staining and flow cytometry in samples originating from the preclinical or clinical setting.

MHC multimer staining has a high sensitivity and specificity for detecting antigen-specific T cells and has been heavily used for investigating epitope spreading and tracking immunotherapy-induced immune responses in both the preclinical and clinical setting in the past. We have also used MHC multimer staining for quality control of T cell receptor-transduced cellular products and in investigational selection of cellular product candidates. Furthermore, MHC multimer staining can be used for testing the effect of repeated freeze/thaw cycles for such a cellular product.

MHC multimer staining utilizes the pMHC-TCR recognition

Common MHC Multimer Staining_pMHC interaction

Antigen-specific T cells recognize specific peptide-MHC (pMHC) complexes on the cell surface and bind to their corresponding T cell receptor (TCR). MHC multimers, which are recombinantly produced pMHC complexes coupled with fluorescent labels are used for detecting antigen-specific T cell responses in fluid samples, e.g. from blood, tumor-infiltrating lymphocytes or cultured cells by flow cytometry. MHC multimer staining are used to detect the presence of antigen-specific T cells and can be combined with our immunophenotyping assays, e.g. to detect the maturation profile of antigen-specific T cells. As IFN-γ-releasing assays such as ELISPOT or intracellular cytokine staining detect only the functional antigen-specific T cells, we highly recommend using MHC multimer staining for achieving validation and a more detailed insight in the antigen-specific T cell population. This is our most exact assay for detection of antigen-specific T cells and requires knowledge about the tissue type of the individual or animal in question.

We ensure the procurement of high-quality MHC monomers and multimers from our distributor Immudex.

For specialized purposes we offer the highly multiplexed dCODE™ Dextramer platform, enabling screening of more than 1000 different antigen-specific T cell responses in parallel. For more information see our dedicated site on this innovative method.

Applications of MHC multimer staining by flow cytometry

  • Detection, enumeration and isolation of antigen-specific T cells in fluid cell samples (PBMCs, tumor-infiltrating lymphocytes or from cell cultures)
  • Antigen-specific immune monitoring of patients in clinical trials for immunotherapy and vaccine development
  • Quality control/quality assurance and selection of cellular products expressing TCRs with certain T cell specificities
  • In vitro experiments in discovery and preclinical development phases for hit-lead optimization
  • Validation of neo-antigen-specific T cell responses in samples from animal models or human patients
  • Investigation of epitope mapping for selection of antigens and epitope spreading for detection of treatment-induced antigen-specific T cell responses.


We offer MHC multimer staining analyses for human and murine samples.

Learn more about

Learn how the MHC multimer dCODE dextramer platform can be used within both immune-oncology and the field of autoimmune diseases, and how ImmuMap offer analyses using DNA barcoded MHC complexes for human samples.