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Cryopreservation of MHC multimers: Recommendations for quality assurance in detection of antigen specific T cells
Author(s) -
Hadrup Sine Reker,
Maurer Dominik,
Laske Karoline,
Frøsig Thomas Mørch,
Andersen Sofie Ramskov,
Britten Cedrik M.,
van der Burg Sjoerd H.,
Walter Steffen,
Gouttefangeas Cécile
Publication year - 2015
Publication title -
cytometry part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.316
H-Index - 90
eISSN - 1552-4930
pISSN - 1552-4922
DOI - 10.1002/cyto.a.22575
Subject(s) - major histocompatibility complex , mhc class i , cryoprotectant , flow cytometry , epitope , cryopreservation , antigen , chemistry , microbiology and biotechnology , biology , computational biology , immunology , embryo
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. © 2014 2014 The Authors. Published by Wiley Periodicals, Inc.