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Rapid and Reliable Custom Tetramer Generation for T Cell Staining

Published by Beata Boczkowska, Ph.D. on

Rapid and Reliable Custom Tetramer Generation for T Cell Staining

The potential for personalized cancer treatment is fueling the need to identify T cell responses against neoantigens and other cancer-specific epitopes for the success of immunotherapy. Continuous advancements of epitope discovery prediction technology is leading to precise identification of antigen-specific T cells, playing a central role in monitoring immune responses to infection and cancer immunotherapies. Hence, the understanding of major histocompatibility complex class (MHC) molecules and peptides interaction within the immune system is fundamental for developing treatments in diseases like cancer and the creation of innovative vaccines.

Fundamentally, in vivo interaction between processed antigen loaded on MHC molecules is important communication for the adaptive immune response to alert against foreign antigens or cancerous cells. MHC I and II molecules loaded with foreign antigens or cancerous fragments are of great interest to the activation of the adaptive immune response. In vivo, peptide exchange reactions are not required for presentation of antigens by MHC molecules because they bind degraded antigens during assembly in the ER. However, peptide exchange reactions play an important role in the assembly of MHC molecules in vitro. It becomes essential to consider the allelic variation and peptide binding when utilizing MHC molecules for T cell detection ex vivo. It has been shown that immunogenic peptides tend to interact with their restricting MHC molecule. Thus, having the capability to assess the binding affinity of an in vitro interaction between peptide and MHC I is highly valued. Consequently, in vitro detection of T cells is challenging because the HLA type and peptide specificity requirements are needed for quantitative analysis of T cell frequency.

MBLI QuickSwitch™ kit technology aims to replicate the immune response by mimicking peptide exchange on an MHC molecule. In vivo, for the T cell epitope to be immunogenic, it must bind to a compatible MHC molecule and remain bound long enough to be presented to and recognized by T cells, thereby eliciting an immune response. Therefore, a strong binding affinity of the MHC/peptide sequence may indicate potential strong immune interactions of antigen-specific T cells. To test this crucial peptide-MHC partnership in vitro, various technologies are available with varying degrees of advantages and disadvantages. The benefits of MBLI peptide-MHC exchange technology is the simplicity and easy to follow method that eliminates the need for complex techniques like UV cleavage. This breakthrough makes the production and evaluation of custom tetramers accessible to labs of all sizes and skill levels. QuickSwitch™ MHC tetramer comes loaded with a placeholder peptide that can be exchanged for a peptide of interest in a process that requires hours instead of weeks. The platform also allows quantification of the placeholder and peptide of interest, which supports evaluation of MHC-peptide combinations for use in tetramer assays and empowers functional screening of immunogenic peptides from infectious agents and cancer neoantigens—invaluable in vaccine development.

Would you like to learn more about how to assess immunogenicity and produce custom tetramers in a matter of hours at your facility? Download this video, where Yuri Poluektov, Ph.D., discusses the use and utility of this versatile tool for creating custom tetramers and screening peptides.

DOWNLOAD THE VIDEO


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