A Reproducible Sequence-Level Strategy to Enhance Peptide Immunogenicity While Preserving Wild-Type Epitope...

Area of Science:

• Immunology
• Peptide Chemistry

Background:

• Short peptide epitopes are crucial for mechanistic studies but often exhibit low immunogenicity.
• Lack of commercially available antibodies against short peptides impedes rapid antibody generation.

Purpose of the Study:

• To develop a reproducible workflow for enhancing peptide immunogenicity.
• To generate epitope-directed antibodies when commercial reagents are unavailable.

Main Methods:

• A sequence-level workflow integrating cross-species/structural triage and MHC-I/II prioritization.
• Conservative heteroclitic-style substitutions to enhance predicted MHC affinity.
• Conjugation of optimized peptides to KLH for antibody induction in mice.

Main Results:

• Optimized peptides demonstrated improved MHC-binding predictions.
• Enhanced antibody titers and isotype maturation (increased IgG1) were observed.
• Generated antibodies retained binding to wild-type sequences, confirming cross-recognition.

Conclusions:

• Strategic, minimal sequence edits significantly enhance peptide immunogenicity.
• Preservation of native epitope recognition is achievable with this method.
• This workflow offers a low-barrier approach for generating custom peptide antibodies.