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High throughput functional epitope mapping: revisiting phage display platform to scan target antigen surface.

Gertrudis Rojas1, Yaima Tundidor, Yanelys Cabrera Infante

  • 1a Systems Biology Department ; Center of Molecular Immunology ; La Habana , Cuba.

Mabs
|December 9, 2014
PubMed
Summary

Developing a robust epitope mapping platform using phage display technology allows for precise identification of antibody binding sites. This method enhances antibody engineering by defining unique functional identities through high-throughput antigen scanning.

Keywords:
Abs, antibodiesAg, antigenEGFEGF receptorEGF, epidermal growth factorEGFR, EGF receptorELISA, enzyme-linked immunosorbent assayIL-2IL-2, interleukin-2PCR, polymerase chain reactionVEGFVEGF, vascular endothelial growth factoraa, amino acidepitope mappinglibrarymAb, monoclonal Abphage displaysite-directed mutagenesis

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Immunology

Background:

  • Antibody engineering requires precise epitope mapping for functional characterization.
  • High-throughput epitope determination is a significant technical challenge.

Purpose of the Study:

  • To develop a robust epitope mapping platform utilizing phage display technology.
  • To overcome challenges in high-throughput fine specificity determination.

Main Methods:

  • Revisiting and adapting filamentous phage display technology.
  • Manipulating target-derived molecules (antigens, domains, fragments) on phages.
  • Employing single, multiple, and combinatorial mutagenesis for comprehensive antigen surface scanning.

Main Results:

  • Successful identification of residue clusters forming epitopes.
  • Delineation of functional epitope pictures within their natural context.
  • Demonstrated versatility in mapping antibodies against interleukin-2, EGF, its receptor, and VEGF.

Conclusions:

  • Phage display, combined with mutagenesis, offers a versatile and high-throughput solution for epitope mapping.
  • This approach accurately defines antibody functional identity by precisely mapping epitopes.
  • The platform is applicable to various antibody-antigen systems, including therapeutic targets.