Cryopreservation of MHC multimers: Recommendations for quality assurance in detection of antigen specific T cells

Cytometry A. 2015 Jan;87(1):37-48. doi: 10.1002/cyto.a.22575. Epub 2014 Oct 8.


Fluorescence-labeled peptide-MHC class I multimers serve as ideal tools for the detection of antigen-specific T cells by flow cytometry, enabling functional and phenotypical characterization of specific T cells at the single cell level. While this technique offers a number of unique advantages, MHC multimer reagents can be difficult to handle in terms of stability and quality assurance. The stability of a given fluorescence-labeled MHC multimer complex depends on both the stability of the peptide-MHC complex itself and the stability of the fluorochrome. Consequently, stability is difficult to predict and long-term storage is generally not recommended. We investigated here the possibility of cryopreserving MHC multimers, both in-house produced and commercially available, using a wide range of peptide-MHC class I multimers comprising virus and cancer-associated epitopes of different affinities presented by various HLA-class I molecules. Cryopreservation of MHC multimers was feasible for at least 6 months, when they were dissolved in buffer containing 5-16% glycerol (v/v) and 0.5% serum albumin (w/v). The addition of cryoprotectants was tolerated across three different T-cell staining protocols for all fluorescence labels tested (PE, APC, PE-Cy7 and Quantum dots). We propose cryopreservation as an easily implementable method for stable storage of MHC multimers and recommend the use of cryopreservation in long-term immunomonitoring projects, thereby eliminating the variability introduced by different batches and inconsistent stability.

Keywords: MHC multimer; cryopreservation; cryoprotectant; glycerol in T cell staining; quality assurance; recommendations for MHC multimer storage.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cryopreservation*
  • Cryoprotective Agents / chemistry
  • Flow Cytometry / standards*
  • Fluorescent Dyes / chemistry
  • Histocompatibility Antigens Class I / chemistry*
  • Humans
  • Indicators and Reagents / standards*
  • Peptides / chemistry*
  • Protein Multimerization
  • Quality Control
  • Quantum Dots / chemistry
  • Reproducibility of Results
  • Staining and Labeling / standards*
  • T-Lymphocytes / immunology
  • T-Lymphocytes / pathology


  • Cryoprotective Agents
  • Fluorescent Dyes
  • Histocompatibility Antigens Class I
  • Indicators and Reagents
  • Peptides