Liquid chromatography-mass spectrometry (LC/MS) is a very powerful analytical tool in the field of proteomics and an important technique for the detection of MHC-associated peptides. Creative BioMart offers services for the analysis of immunogenicity-related peptides presented on the MHC, known as MHC-associated peptide proteomics (MAPP), which can be applied to search for antigens presented on the MHC for drug development.

MHC-associated Peptide Proteomics

Primary immune responses to antigens are initiated by antigen-presenting cells such as dendritic cells (DCs), which deliver protein-derived peptide fragments on the surface of these cells, with MHC I-associated antigens presenting peptides to CD8+ T cells and MHC II-associated antigens presenting to CD4+ T cells. Therefore, the identification of these relevant peptides via MHC presentation creates great potential for the generation of new therapeutic agents, vaccines and diagnostic agents.

The use of LC/MS to study the composition and kinetics of peptides presented by MHC molecules is important for the discovery of T cell targets to combat cancer as well as epidemic pathogens. We can compare peptides from MHC-presented virally infected and tumor cells to equivalent healthy cells using MHC-associated peptide proteomics assays. Using powerful MS analysis, we can identify antigens that can serve as therapeutic targets, and in combination with in vitro T-cell assays can confirm whether peptides isolated and identified from MHC molecules contain epitopes that can trigger T-cell responses.

MAPP Assay

MAPP assays identify MHC-associated peptides from across the proteome, and the identified regions can be used as potential T cell epitopes or immune targets. We offer a one-stop service for the assay as follows.

Fig.1 Service flow of MAPP assay

• Prepare DCs produced from healthy donors and loaded with test biotherapeutic proteins.
• DCs take up, process and present peptides from proteins via MHC-related molecules.
• Lyse and immunoprecipitate DCs and isolate relevant peptides from them.
• Identify MHC-associated peptides by LC/MS, mapping peptides to biotherapeutic protein sequences.
• Predict the risk of immunogenicity of identified peptides, aided by a T-cell assay.

Application Areas

Antigen Discovery and Validation

We can identify tumor-associated, pathogen-associated antigens based on comparison with healthy tissue, synthesize any variable-presentation protein peptide to check T-cell activation, and discover and validate new targets for immunotherapy.

Vaccine Design

Identify potential antigens for vaccine design. Relevant antigens are expressed in human cell lines and analyzed for comparative presentation of peptides, and the unique viral peptides identified are synthesized and used in an in vitro T cell assay to assess T cell responses.

Diagnosis Development

Identify relevant T cell epitopes from the whole proteome and antigens and use in assays designed to monitor effective T cell responses in patient isolates to diagnose disease outcomes.

Creative BioMart tailors a specific service plan for each client's unique project to ensure that your desired results are met or exceeded. Our experienced, professional, reliable and hands-on team is focused on moving your project forward until results are achieved, and is committed to being a professional and attentive partner in providing quality services.

If you would like more information about our services, a project quote or professional consultation, please contact us.

Reference

• Nobuo S, et al. "MHC-associated peptide proteomics enabling highly sensitive detection of immunogenic sequences for the development of therapeutic antibodies with low immunogenicity." Mabs (2018),10(8):1168-1181.