Journal
CYTOMETRY PART A
Volume 87A, Issue 1, Pages 37-48Publisher
WILEY
DOI: 10.1002/cyto.a.22575
Keywords
MHC multimer; cryopreservation; cryoprotectant; recommendations for MHC multimer storage; quality assurance; glycerol in T cell staining
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Funding
- Deutsche Forschungsgemeinschaft [SFB 685/Z5]
- Danish Council for Strategic Research
- Danish Cancer Society
- Carlsberg Foundation
- Jochum Foundation
- Wallace H. Coulter Foundation (WCF, Miami, Florida)
- The Danish Cancer Society [R72-A4531] Funding Source: researchfish
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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. (c) 2014 International Society for Advancement of Cytometry
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