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A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits
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Published on: February 16, 2014

Phage display and selections on cells.

Klervi Even-Desrumeaux1, Patrick Chames

  • 1INSERM U624, Marseille, France.

Methods in Molecular Biology (Clifton, N.J.)
|August 22, 2012
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Summary
This summary is machine-generated.

This study introduces a novel cell surface phage display panning method. This technique enables the discovery of antibodies targeting cell surface receptors, even when the antigen is unknown.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Immunology

Background:

  • Traditional phage display panning requires purified antigens immobilized on solid phases.
  • This limitation restricts its application for cell surface targets or unknown antigens.
  • Developing methods for cell surface target identification is crucial for antibody discovery.

Purpose of the Study:

  • To describe a novel phage display panning strategy for targeting cell surface receptors.
  • To detail a protocol including a depletion step for enhanced specificity.
  • To provide a comprehensive guide for antibody discovery against cell surface antigens.

Main Methods:

  • Production of a phage library.
  • Phage display panning on cell surface receptors with a depletion step.
  • Elution of bound phages.
  • Screening of soluble antibodies using ELISA and flow cytometry.

Main Results:

  • The described protocol allows for the selection of phages displaying antibodies against cell surface receptors.
  • The method includes steps for depletion, selection, elution, and screening.
  • Analysis of soluble antibodies confirms their binding specificity via ELISA and flow cytometry.

Conclusions:

  • The developed cell surface phage display panning strategy is effective for identifying antibodies against cell surface targets.
  • This method overcomes limitations of traditional panning by enabling selection directly on cells.
  • The protocol offers a valuable tool for antibody discovery in various biological and therapeutic applications.