Creative BioMart offers testing services to measure the stability assessment of your peptide in complex with MHC molecules, which is an aspect of immunogenicity assessment. The stability of the MHC/peptide complex is also critical for determining epitope immunogenicity. The stability of any given peptide-MHC complex can be determined by monitoring the dissociation of the immune complex at 37°C and calculating the half-life. Our service can utilize custom MHC molecules for testing.

MHC/Epitope Stability and Immunogenicity Assessment

In the 1990s, it was recognized that the immunogenicity of peptides bound to MHC class I molecules depends on the stability of the MHC-peptide complex. For example, efficient presentation of peptide-MHC class I (pMHC-I) complexes to immune T cells benefit from stable peptide-MHC-I interactions. Therefore, the stability of MHC/peptide complexes is probably the most important parameter in determining epitope immunogenicity and must be assessed to reduce false positive epitope binders. Assessment of peptide-MHC complex stability is an important parameter when screening immunogenic peptides. The stability of any given peptide-MHC complex can be determined by monitoring the dissociation of the immune complex at 37°C and calculating the half-life. Studies have shown that the half-life of the peptide-MHC I complex correlates better with immunogenicity than the affinity of the corresponding peptide for MHC.

Fig.1 Stability is a better indicator of immunogenicity than affinity. (Mikkel Harndahl et al., 2012)

Featured MHC/Peptide Stability Assay

This service forms an important part of evaluating the immunogenicity of MHC I and II /epitope complexes, and single stability or affinity assessment may yield inaccurate and false-positive results.

Creative BioMart stability assays are achieved by measuring the half-life of MHC/peptide complexes under pressure-inducing conditions such as pH or temperature changes or urea denaturation. We utilized custom reference peptides for quantification and calculated stability relative to the included reference peptides. From this, the half-life of each MHC/peptide complex can be determined. Reference peptides were included in all assays, and data generated from test epitopes were expressed as a percentage of reference peptides. In general, epitopes with Kd below 100 nM are considered potentially immunogenic. This stability assay is also suitable for high-throughput screening and does not require any peptide modification. Our suppliers can provide monomeric or tetrameric protein products for testing to complete the understanding of the potential immunogenicity of the molecule of interest.

Fig.2 Featured MHC/Peptide Stability Assay Services.

Advantages of Creative BioMart

• Choice of multiple pressure-induced conditions, including pH, temperature changes, urea denaturation, etc.;
• Quantification by custom reference peptides;
• Comprehensive analysis of optional affinity testing services;

Creative BioMart offers high quality MHC-peptide complex stability testing services as part of immunogenicity assessment. Assessment of stability is achieved by alignment with reference peptides. Our stability testing service can be combined with the results of affinity testing to help researchers evaluate immunogenicity more comprehensively. Our experimental platform enables high-quality analytical testing. If you are interested in our services, please contact us for cooperation.

References

• Harndahl, Mikkel, et al. "Peptide‐MHC class I stability is a better predictor than peptide affinity of CTL immunogenicity." European journal of immunology 42.6 (2012): 1405-1416.
• Kenneth L. Rock, et al. "Present Yourself! By MHC Class I and MHC Class II Molecules" (2016). Trends Immunol. 37(11): 724 –737.