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Sheep monoclonal antibody fragments generated using a phage display system.

Y Li1, J Kilpatrick, G C Whitelam

  • 1Department of Biology, University of Leicester, University Road, Leicester, UK. YL4@leicester.ac.uk

Journal of Immunological Methods
|March 4, 2000
PubMed
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Researchers developed a novel phage display method to create recombinant sheep antibody fragments. This technique offers a valuable tool for sheep immunology research and general biorecognition applications.

Area of Science:

  • Immunology
  • Biotechnology
  • Molecular Biology

Background:

  • Monoclonal sheep antibodies hold significant promise for biomedical, veterinary, and agricultural applications.
  • Conventional methods for generating sheep monoclonal antibodies, such as hybridoma technology, are not routine and can be complex.

Purpose of the Study:

  • To establish an efficient method for generating recombinant sheep antibody fragments from immunized animals.
  • To develop a valuable tool for studying sheep immunology and for general biorecognition.

Main Methods:

  • Utilized a modified phage display system to generate recombinant antibody fragments.
  • Amplified immunoglobulin V gene repertoires from the spleens of sheep immunized with human serum albumin and conalbumin.
  • Constructed a phage display single-chain variable fragment (scFv) library using an efficient two-step cloning method.

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Main Results:

  • Successfully isolated and characterized 14 different single-chain Fv (scFv) fragments.
  • Sequence analysis confirmed typical ovine immunoglobulin characteristics, identifying thirteen Vlambda and 11 VH genes.
  • Produced soluble monomeric scFvs in Escherichia coli and measured affinities using surface plasmon resonance, observing affinities typical of a secondary immune response.

Conclusions:

  • The described method provides a robust approach for generating recombinant sheep antibody fragments.
  • This technique is valuable for advancing sheep immunology research and facilitating biorecognition studies.
  • The generated scFvs exhibit characteristics suitable for various biotechnological applications.