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Research

MHC Tetramer Production Services

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Our core provides customized MHC/peptide tetramers for identification of antigen specific T lymphocytes by flow cytometry. Tetramers and/or their derivatives are provided for research purposes only.

Choices of fluorescent labels: R-phycoerythrin (PE) or allophycocyanin (APC)

Standard product size: 100μg of MHC/peptide complex protein, which is sufficient for 400 tests (PE labeled) or 250 tests (APC labeled).

Other product size: We may split standard vials into smaller aliquots upon request; aliquot size can be as low as 25μg each.

Monomers: Monomeric soluble MHC/peptide complex, biotinylated and non-biotinylated, is available upon request.

Clinical Research Grade Reagents: We provide Class I tetramers for certain clinical researches.

Special Reagents:

  • CD8 binding-deficient MHC Class I tetramers:

    Containing mutations in the MHC α3 domain that ablate CD8 binding; can be used to quantitate and to sort CD8-independent T cells.
     
  • Chimeric Tetramer:

    Containing MHC Human/Mouse chimeric heavy-chain.
     
  • MHC Monomers for generating TCR-like antibodies:

    Developing high-affinity soluble antibody molecules endowed with a TCR-like specificity toward tumor epitopes, termed TCR-like antibodies, recently became a hot topic for cancer immunotherapy studies.

    Major histocompatibility complex (MHC) class I presentation of peptides to T cell receptors (TCR) on T lymphocytes is the key in the immune response. Because of their unique specificity, MHC-peptide complexes, which can be recognized by recombinant T-cell receptors (TCRs), are desirable candidates for novel immunotherapeutic approaches. However, most TCRs produced previously have affinities that are too low for target detection under normal assay conditions, and limited stability (due to their generation in a single-chain form). Therefore, the TCR-like antibodies are being developed as a new immunotherapeutic class for targeting tumor cells and mediating their specific killing, by constructing recombinant immunotoxins, fusions with cytokine molecules, or for bi-specific antibody therapy. Meanwhile, these antibodies are also valuable research reagents enabling the study of class I peptide ligand presentation and TCR-peptide-MHC interactions.

    Our biotinylated or unbiotinylated monomers can be used to generate TCR-like antibodies, as well as in the affinity characterization approaches.
     
  • MHC Class I monomers and tetramers designed for ligand exchange:

    When trying to analyze multi epitopes of an antigen, generating individual tetramers can be very time consuming and less practical. The MHC monomers specially designed for ligand exchange can address this issue well. Conditional MHC ligands form stable complexes with MHC molecules but degrade on command by exposure to UV light; Then generate ‘Empty MHC molecules’ and load with different peptide ligands. In addition, because ligand exchange allows the direct use of MHC multimers that are generated in a procedure about an hour without further purification steps, the technology is also well-suited to study immune responses against low affinity ligands such as cancer- or autoimmune disease-associated epitopes.
     
  • SARS-CoV-2/COVID-19 Tetramer Reagents: 

    We produce a set of SARS-CoV-2/COVID-19 tetramer reagents based on published work. Epitopes were previously identified in SARS and are 100 percent conserved in SARS-CoV-2.
     
    • HLA-A*02:01/SARS-CoV-2S.940~948 (ALNTLVKQL)
    • HLA-A*02:01/SARS-CoV-2S.958~966 (VLNDILSRL)
    • HLA-A*02:01/SARS-CoV-2S.978~986 (LITGRLQSL)
    • HLA-A*02:01/SARS-CoV-2S.1167~1175 (RLNEVAKNL)
    • HLA-A*02:01/SARS-CoV-2S.1192~1200 (NLNESLIDL)
    • HLA-A*02:01/SARS-CoV-2S.1220~1228 (FIAGLIAIV)
    • HLA-A*02:01/SARS-CoV-2S.269~277 (YLQPRTFLL)
    • HLA-A*02:01/SARS-CoV-2N.222~230 (LLLDRLNQL)
    • HLA-A*02:01/SARS-CoV-2M.61~70 (TLACFVLAAV)
    • HLA-A*02:01/SARS-CoV-2S.816~826 (SFIEDLLFNKV)
    • HLA-A*03:01/SARS-CoV-2S.938~947 (LSSTASALGK)
    • HLA-A*24:02/SARS-CoV-2S.489~497 (YFPLQSYGF)
    • H-2D(b)/SARS-CoV-2S.538~546 (CVNFNFNGL)
    • H-2D(b)/SARS-CoV-2S.350~358 (VYAWNRKRI)
    • H-2D(b)/SARS-CoV-2S.366~374 (SVLYNSASF)
    • H-2D(b)/SARS-CoV-2S.444~452 (KVGGNYNYL)
    • Mamu-A1*00101/SARS-CoV-2S.712~720 (IAIPTNFTI)
    • Mamu-A1*00101/SARS-CoV-2S.1055~1063 (SAPHGVVFL)
    • Mamu-A1*00101/SARS-CoV-2S.489~497 (YFPLQSYGF)
    • Mamu-A1*01101/SARS-CoV-2N.322~331 (MEVTPSGTWL)
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Available MHC Alleles

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Refer to Table 1 for the current available MHC alleles.

If the requested MHC allele is not in the table or if there is other requirements, such as a specific b2-microglobulin is needed to reconstitute class I of an unusual species, the requestor must supply the cDNA. We will perform the subcloning to the appropriate vector for protein expression and additional charges may apply.

All expression plasmids containing MHC alleles will be retained at our facility and will be used for future production and distribution of tetramer reagents.

All expression plasmids containing MHC alleles will be retained at our facility and will be used for future production and distribution of tetramer reagents.

Pre-Made Common Tetramers

We offer some common class I reagents from a pre-made stock while accepting custom synthesis requests. Please refer to the list below.

Peptide

Requestors incur the cost of peptide that is needed in the tetramer production. You can either provide your own peptide or order from us with an additional cost. If provide your own peptide, please send 10mg of it to us. HPLC chromatograph and mass spectrophotometric analysis for the peptide will also be required.

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