Quantitative Assessment of Antigen Peptide-MHC Binding Affinity and High-Throughput ELISA Screening Platform for Population Adaptability

This technical platform offers precise evaluation services for the binding affinity between antigenic peptides and HLA-I molecules, enabling efficient screening of antigenic epitopes with HLA-I-restricted presentation potential. The technological system has been maturely applied in the following domains: 1) Development of immunotherapy targets for tumors (e.g., tumor neoantigen screening), 2) Personalized tumor vaccine preparation (including custom synthesis of MHC-I multimers), and 3) Vaccine research for infectious diseases (e.g., monitoring of virus-specific T cell immune responses).

Immunity/InflammaAntigen peptide MHC binding affinity ELISAMHC
  • Recent Advances
  • Product Information
Recent Advances
MHC Antigen Peptide Multimer Technology
Technical Platform Introduction
Our platform specializes in accurately evaluating the binding affinity of antigenic peptides to HLA class I molecules, efficiently identifying antigenic epitopes with HLA class I presentation potential. This technology has been successfully applied in oncology immunotherapy target discovery, personalized cancer vaccine development, infectious disease vaccine research, and more.
Core Technical Principles
In the presence of a target antigenic peptide, the HLA class I heavy chain and β2-microglobulin (B2M) assemble into a trimeric complex via non-covalent interactions, accompanied by specific conformational changes. Using an anti-B2M-HRP antibody as a detection probe, and with a streptavidin-coated plate or anti-HLA monoclonal antibody solid-phase system, the complex is captured via a sandwich ELISA method. By quantifying the chromogenic reaction intensity at 450 nm, a standard curve correlating antigenic peptide concentration with binding affinity is established for semi-quantitative assessment. Peptides with insufficient binding affinity or unable to stabilize the complex fail to anchor to the solid-phase carrier, leading to negative ELISA results post-washing. The system ensures specificity through dual quality controls (blank and positive peptide controls) and achieves a detection limit as low as 1 nM.
Application Values
  • Tumor Neoantigen Screening: Quantify HLA class I presentation efficiency of tumor mutation-derived peptides to predict immunogenicity, aiding in the selection of personalized tumor-specific neoantigens. Combined with patient HLA typing data, it helps analyze antigen presentation defects in tumor immune evasion.
  • Tumor Vaccine Target Evaluation: Construct a high-throughput screening platform covering common HLA alleles to assess peptide library cross-reactivity and optimize epitope affinity, guiding rational design and immunogenicity enhancement of tumor peptide vaccines.
  • TCR-T Preclinical Target Validation: Establish clinical-grade MHC multimer production processes based on validated antigenic peptide-MHC complexes, supporting the entire workflow from single-cell sequencing data to functional TCR screening, accelerating in vitro expansion and affinity maturation of antigen-specific T cells.
  • Viral Antigen Peptide Screening: Monitor antigenic drift in viral variants to build HLA-restricted epitope databases, providing critical epitope information for broadly protective vaccine design and assessing the impact of viral escape mutations on T cell responses.
  • Vaccine Immunity Assessment: Evaluate antigen-specific T cell frequency and functional phenotypes post-vaccination to establish an immune response quality assessment system, linking in vitro data to in vivo protection and guiding vaccine dosing and scheduling optimization.
  • Autoimmune Antigen Screening: Analyze abnormal antigen presentation spectra of HLA risk alleles linked to autoimmune diseases, identifying pathogenic T cell-specific self-antigen epitopes for disease classification and targeted immunotherapy.
  • Organ Transplant Antigen Selection: Develop HLA mismatched antigen epitope prediction algorithms for donors and recipients, establish an indirect recognition epitope library related to transplant rejection, and support strategies for immunologically inert donor organ modification and recipient-specific desensitization.
Assay Formats
Our platform offers three standardized assay formats tailored for different research needs:
  1. Antigen Peptide Screening Mode: Evaluate high-throughput binding of peptides to a single HLA allele. This mode rapidly identifies HLA-binding tumor neoantigens, viral epitopes, and autoimmunity-related antigens. Services include peptide libraries (≥50 sequences), standard HLA monomers, and assay kits, with results within 72 hours. Ideal for oncology target libraries, viral escape monitoring, and autoimmune disease research.
  2. HLA Compatibility Mode: Map antigenic peptides to HLA alleles across a broad population spectrum. Using microarray chip technology to test 48 common alleles, this mode calculates epitope recognition rates in specific HLA-positive populations. It supports vaccine design and guides region-specific or personalized immunotherapy strategies.
  3. Complex Validation Mode: Validate and optimize pMHC complex production processes described in literature or patents. Services include:
  • HLA heavy chain/B2M mutant binding assays.
  • Folding buffer optimization via pH and salt gradient screening.
  • Purification tag efficiency evaluation.
  • Stability testing at 4℃ and 37℃.
Comprehensive quality control through SDS-PAGE, DLS, and functional assays ensures the complexes meet quality standards for TCR antibody development, tetramer staining reagent production, and CAR-T cell expansion.
Related Products:
Disease Category Product Name Antigen Sequence MHC Position Product Number
EBV HLA-A*0201/YLELLVWRL-PE Labelled Tetramer EBV.LMP1 YLELLVWRL HLA-A*0201 125-133 UA089001
EBV HLA-A*0201/YLQQNWTL-PE Labelled Tetramer EBV.LMP1 YLQQNWTL HLA-A*0201 159-167 UA089003
EBV H-2Db(b)/RAHY-NIVTF-PE Labelled Tetramer HPV16.E7 RAHYNIVTF H-2Db 49-57 UA089002
HPV H-2K(b)/EVYDFA-FRQL-PE Labelled Tetramer HPV16.E6 EVYDFARDL H-2Kb 48-57 UA089004
HPV HLA-A*0201/KLP-DLCTL-PE Labelled Tetramer HPV18.E6 KLPDCTL HLA-A*0201 13-21 UA089005
HPV HLA-A*0201/KLTNT-GLYQL-PE Labelled Tetramer HPV18.E6 KLTNTGLYNL HLA-A*0201 92-101 UA089006
HPV HLA-A*0201/TLODIVIHL-PE Labelled Tetramer HPV18.E7 TLODIVIHL HLA-A*0201 7~15 UA089007
HPV HLA-A*0201/QFLNTL-FV-PE Labelled Tetramer HPV18.E7 QFLNTLFSV HLA-A*0201 88-97 UA089008
HPV HLA-A*1101/GVNHQLPAR-PE Labelled Tetramer HPV18.E7 GVNHQLPAR HLA-A*1101 43-52 UA089009
Influenza A Virus H-2D(b)/ASNENMETM-PE Labelled Tetramer Flu.NP ASNENMETM H-2Db 366-374 UA089010
Influenza A Virus H-2K(d)/TYQR-TRALY-PE Labelled Tetramer Flu.NP TYQRTRALY H-2Kd 147-155 UA089011
Influenza A Virus H-2D(b)/ASNEN-MDTM-PE Labelled Tetramer Flu.NP ASNENMDTM H-2Db 366-374 UA089012
LCMV H-2D(b)/KAVYNFATM-PE Labelled Tetramer GP 33 KAVYNFATM H-2Db 33-41 UA089013
LCMV H-2D(b)/FQPGQGFVK-PE Labelled Tetramer LCMV NP FQPGQGFVK H-2Db 396-404 UA089014
Tumor-related HLA-A*1101/VVGADGVK-PE Labelled Tetramer KRAS VVGADGVK HLA-A*1101 7~16 UA089015
Tumor-related HLA-A*1101/VVGAGVGK-PE Labelled Tetramer KRAS VVGAGVGK HLA-A*1101 7~16 UA089016
Tumor-related HLA-A*0201/KLVVGAGV-PE Labelled Tetramer KRAS KLVVGAGV HLA-A*0201 5~14 UA089017
Tumor-related HLA-A*0201/SLLMWITQC-PE Labelled Tetramer NY-ESO1 SLLMWITQC HLA-A*0201 157-165 UA089018
Melanoma HLA-A*0201/LMWITQCFL-PE Labelled Tetramer NY-ESO2 LMWITQCFL HLA-A*0201 159-167 UA089019
Melanoma H-2Db(b)/MMFPNA-P1-PE Labelled Tetramer WT1 RMFPNAPL H-2Db 126-134 UA089020
Melanoma HLA-A*0201/CMTWV-PE Labelled Tetramer WT2 CMTWVNMDM HLA-A*0201 235-243 UA089021
Melanoma HLA-A*1101/KTCQRKSF-PE Labelled Tetramer WT3 KTCQRKSF HLA-A*1101 386-394 UA089022
Ovarian Cancer H-2K(b)/SINFEKL-PE Labelled Tetramer OVA SINFEKL H-2Kb 257-264 UA089023

This article is reviewed and published by the technical expert team of UA

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