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Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
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Related Experiment Video

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Laboratory Scale Production and Purification of a Therapeutic Antibody
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Recombinant antibody fragment production.

H Ma1, R O'Kennedy1

  • 1Biomedical Diagnostics Institute, Dublin City University, Dublin 9, Ireland.

Methods (San Diego, Calif.)
|November 23, 2016
PubMed
Summary

This study details constructing a recombinant antibody library using single-chain variable antibody fragments (scFv). It showcases an efficient screening and purification method for these valuable therapeutic and diagnostic tools.

Keywords:
PanningPurificationRecombinant antibodyscFv

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Area of Science:

  • Biotechnology
  • Immunology
  • Molecular Biology

Background:

  • Recombinant antibodies offer advantages over conventional ones in therapeutics and diagnostics.
  • Single-chain variable antibody fragments (scFv) are a key recombinant antibody format.
  • Efficient methods for generating and selecting recombinant antibodies are crucial.

Purpose of the Study:

  • To demonstrate the construction of a recombinant antibody library using the scFv format.
  • To present an efficient immunotube-based panning approach for antibody screening.
  • To describe methods for antibody selection and purification.

Main Methods:

  • Construction of a recombinant antibody library using scFv.
  • An immunotube-based, two-day panning approach for antibody screening.
  • Utilisation of Escherichia coli as an expression system.
  • Antibody selection and purification via immobilised metal affinity chromatography (IMAC).

Main Results:

  • Successful generation of a recombinant antibody library.
  • Demonstration of an efficient two-day screening process.
  • Effective purification of recombinant antibodies using IMAC.

Conclusions:

  • The described methods provide an efficient route for constructing and screening recombinant antibody libraries.
  • This approach facilitates the development of scFv antibodies for therapeutic and diagnostic applications.
  • The presented techniques are valuable for antibody engineering and production.