QuickSwitch HLA-A*02:01 Peptide Screening Kit

MBL Life Sciences

Catalog No.: MBL-TS-7300-SR

APPLICATION

The QuickSwitch™ HLA-A*02:01 Peptide Screening Kit utilizes a patented technique for exchanging up to 93 peptides on an MHC class I monomer. Components for quantifying the extent of peptide exchange by flow cytometry are included. The primary intended application of this assay is to screen, in a high throughput fashion, peptides that stably bind to the MHC complex. Usage of this kit is particularly indicated for validating neoantigen sequences obtained by exome high throughput sequencing and prescreened with epitope prediction algorithms.

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SUMMARY AND EXPLANATION

Major histocompatibility complex (MHC)-encoded glycoproteins bind peptide antigens through non-covalent interactions to generate complexes that are displayed on the surface of antigen-presenting cells for recognition by T cells. Peptide-binding site occupancy is necessary for stable assembly of newly synthesized MHC proteins and export from the endoplasmic reticulum. During this stage peptides produced in the cytosol compete for binding to MHC molecules, resulting in extensive peptide exchanges that are regulated by accessory molecules, such as tapasin.¹² The QuickSwitch™ HLA-A*02:01 Peptide Screening Assay is based on the capacity of MHC class I molecules to exchange peptides.

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PRINCIPLE

The kit contains two modules:

1. Biotinylated MHC class I monomer units folded with an irrelevant exchangeable peptide, along with a proprietary Peptide Exchange Factor, for the generation of monomers loaded with specific peptides of interest.
2. A flow cytometry-based sandwich immunoassay containing streptavidin-conjugated magnetic beads to capture MHC class I monomers and a FITC-labeled antibody recognizing the Exiting peptide.

This assay allows for determination of the capacity of test peptides to bind to MHC by quantifying the displacement of the original peptide. Peptide exchanged MHC molecules have multiple in vitro applications including identification of antigen-specific CD8⁺ T cells after tetramerization with fluorochrome-conjugated streptavidin (Note 1).

Product Specifications

• Application: FC
• Reactivity: Human
• Class: Class I
• MHC Allele: A*02:01

Citations for QuickSwitch HLA-A*02:01 Peptide Screening Kit – 2 Found

Peaper, David R; Cresswell, Peter. Regulation of MHC class I assembly and peptide binding. Annual Review Of Cell And Developmental Biology. 24:343-68. 

Mayerhofer, Peter U; Tampé, Robert. Antigen translocation machineries in adaptive immunity and viral immune evasion. Journal Of Molecular Biology. 2015;427(5):1102-18.