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Epitope mapping of antibodies

Several new technologies for analyzing the binding parameters of antibodies have been developed for antibody mapping. First, libraries using bacterial surface display have been employed to screen epitopes for antibodies of therapeutic interest. Second, microfabricated arrays with many millions of synthetic peptides have been developed and used for fine mapping of antibody epitopes. Third, suspension bead arrays have been used to allow for a flexible system for epitope mapping of both monoclonal and polyclonal antibodies.

Key publication

Other selected publications

  • Rockberg J et al., Prediction of antibody response using recombinant human protein fragments as antigen. Protein Sci. (2009)
    PubMed: 19760667 DOI: 10.1002/pro.245

  • Rockberg J et al., Discovery of epitopes for targeting the human epidermal growth factor receptor 2 (HER2) with antibodies. Mol Oncol. (2009)
    PubMed: 19393584 DOI: 10.1016/j.molonc.2009.01.003

  • Hjelm B et al., Exploring epitopes of antibodies toward the human tryptophanyl-tRNA synthetase. N Biotechnol. (2010)
    PubMed: 19913117 DOI: 10.1016/j.nbt.2009.11.001

  • Buus S et al., High-resolution mapping of linear antibody epitopes using ultrahigh-density peptide microarrays. Mol Cell Proteomics. (2012)
    PubMed: 22984286 DOI: 10.1074/mcp.M112.020800

  • Forsström B et al., Proteome-wide epitope mapping of antibodies using ultra-dense peptide arrays. Mol Cell Proteomics. (2014)
    PubMed: 24705123 DOI: 10.1074/mcp.M113.033308

Figure legend: Peptide arrays have been developed covering all human proteins for the analysis of antibody specificity. The technology was based on parallel in situ photolithic synthesis of millions of overlapping peptides. Here, epitopes are shown for antibodies binding to SOD1 and CD4 as determined by these microarrays. Adapted from Forsström et al. (2014).

Key facts

  • Binding epitopes were dissected using suspension bead arrays
  • Microarrays containing millions of peptides were designed and used for binding analysis
  • Recombinant libraries expressed with bacterial surface display were also used for epitope mapping