Why identifying antigen-specific epitopes and their cognate T-cell receptors is important
In immunology, identifying antigen-specific epitopes and their cognate T-cell receptors can yield useful insights into the functioning of T-cells and can provide vital information for vaccine development. dCODE Dextramer® (RiO) reagents are DNA barcoded MHC Dextramer® products compatible with the BD RhapsodyTM Single-Cell Analysis System. Each barcode can be associated with a unique MHC-peptide epitope that is displayed on the dCODE Dextramer®. The PE fluorochrome labelling on the dCODE Dextramer® allows a FACS sorting enrichment.
The dCODE Dextramer® can be used to determine T-cell – antigen specificity at the single-cell level and provides simultaneous information about gene and protein marker expression. The unique barcode for each MHC specificity allows for multiplexing screening, and the multiple MHC-peptide complexes permit high-avidity binding to T-cell receptors. The use of the BD RhapsodyTM Single-Cell Analysis System makes multi-parameter analysis possible.
Results of this study
In this study, Jacobsen K et.al have detected and analysed disease-specific CD4+ cells within thousands of PBMCs by using the two technologies. They simulated PBMCs with two peptide antigens (EBV and tetanus toxoid) and stained them with a panel of dCODE Dextramers®. Then they captured and lysed the stained cells using the BD RhapsodyTM Single-Cell Analysis System. cDNA synthesis of the captured polyadenylated mRNA and the dCODE Dextramer® barcodes was then carried out following PCR amplification. The data was further analysed using SeqGeq™.
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