High Resolution Epitope Mapping and Antibody Cross-Reactivity Analysis

摘要

Antibodies are among the most important life science tools for therapeutics, basic research and diagnostic tests. However, mono- and polyclonal antibodies are often poorly characterized in terms of specificity and cross-reactivity, needing further validation and cross-reactivity analysis. High density peptide microarrays are ideally suited for antibody characterization, enabling the analysis of epitope-antibody interactions with unmet speed and precision. Based on approaches for linear and conformational epitope mappings as well as high resolution epitope substitution scans, we developed a comprehensive toolbox for the in-depth analysis of epitopes and antibody cross-reactions with all kinds of antibodies and isotypes in the most flexible, comprehensive and economic manner. Moreover, we designed a Human Epitome Microarray comprising all linear human B-cell epitopes of the Immune Epitope Database (23,163 linear peptides), complemented by 4,661 epitopes of the most common vaccines. In a three-step approach based on a single assay, the Human Epitome Microarray enables a very detailed cross-reactivity analysis of antibodies including the identification of antibody-specific consensus motifs and database blasting to identify possibly cross-reacting human antigens. The presentation will hence summarize the different high-density peptide microarray approaches for a comprehensive antibody characterization and cross-reactivity analysis.