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A mammalian cell display platform based on scFab transposition.

Jing Chang1, Christoph Rader1, Haiyong Peng1

  • 1Department of Immunology and Microbiology, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology, University of Florida, Jupiter, FL 33458, USA.

Antibody Therapeutics
|July 26, 2023
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Summary

Researchers developed a novel mammalian cell display platform using the PiggyBac transposon system for efficient discovery of high-affinity monoclonal antibodies (mAbs). This system enables antibody engineering for therapeutic applications.

Keywords:
in vitro display technologiesmammalian cell displayrabbit monoclonal antibodiesscFabtransposon

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Area of Science:

  • Biotechnology
  • Immunology
  • Molecular Biology

Background:

  • In vitro display technologies like phage and yeast display are established for antibody discovery.
  • Mammalian cell-based display systems are emerging to address limitations in post-translational modifications.
  • Developing novel platforms is crucial for advancing antibody engineering.

Purpose of the Study:

  • To introduce a new mammalian cell display platform for antibody discovery.
  • To demonstrate the platform's capability in isolating high-affinity monoclonal antibodies (mAbs).
  • To validate the platform for potential antibody humanization and maturation.

Main Methods:

  • Utilized the PiggyBac transposon system for stable integration into HEK293F cells.
  • Generated immune rabbit antibody libraries (~7x10^7 clones) in an scFab format.
  • Employed magnetic-activated and fluorescence-activated cell sorting for antibody selection.

Main Results:

  • Successfully displayed single-chain Fab (scFab) antibodies on HEK293F cell surfaces.
  • Identified mAbs with subnanomolar affinity and cross-reactivity to human and mouse antigens.
  • Demonstrated the platform's efficacy in antibody discovery and optimization.

Conclusions:

  • The PiggyBac transposon-based mammalian cell display is a powerful tool for antibody discovery.
  • This platform facilitates the isolation of high-affinity and cross-reactive antibodies.
  • The system holds potential for further antibody engineering, including humanization and affinity maturation.