As research experience accumulating in the past couple of years and the techniques developed by our excellent research team in epitope mapping, Creative Biolabs offers the unique MHC-peptide binding assay service for our clients.
This technique is a novel approach to rapidly identify T cell epitopes with short turnaround time. It contains three major steps: the detection of MHC-peptide binding assays, followed by rate assays and cell assays to validate the high MHC-affinity peptides. Peptides used for this assay can be provided by clients or synthesized by our peptide library platform.
Fig 1. The workflow of MHC-Peptide Binding Assay.
Binding assay: The binding of each peptide with MHC molecules aims to measure the capability of candidate peptides to stabilize the structure of MHC-peptide complex, which is reflected by the distinct signal. Only when the peptides could bind to MHC and stabilize the conformation will the signal be released. Each candidate peptide achieves a score relative to the positive control peptide (known T cell epitope). And based on corresponding analysis of signal percentage, the lead selections will be chosen for the next step study.
Rate assay: This assay is followed by the MHC-peptide binding assay and targets for quantitative determination of the binding abilities of the selected peptides. Peptides that are presented with enough length have the greater chance to be recognized by T cells. The on-or/and off-rates for the selected peptides are measured at different time points. Comprehensive analysis and achieve a score for the individual peptide. The higher score represents a better T cell epitope candidate.
Validation assay: The strategies above will cut down the number of candidate epitopes for further research. Then cell assays could be committed to detecting whether these selected peptides have the property to evoke relevant T cell responses. Detection of the T cell responses induced by immunogenic peptides keeps two major approaches: determine the proliferation of T cells or the cytokine-secreting by T cells in the presence of potentially immunogenic peptides. T cell proliferation assays are performed by combining with the MHC multimers (tetramers, Pentamers, dextramers) techniques which are detected by flow cytometer. The cytokine secreting of T cells is committed with the ultrasensitive ELISPOT technique to accurately reflect the antigen-specific T cell responses.
Creative Biolabs has years of experience in offering our customer epitope mapping techniques and we are happy to provide the advanced technical support to help our clients find the T cell epitope. If you are interested in our service, please contact us for more information and a detailed quote.