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Related Experiment Videos

Critical epitopes in transmissible gastroenteritis virus neutralization.

G Jiménez, I Correa, M P Melgosa

    Journal of Virology
    |October 1, 1986
    PubMed
    Summary

    Monoclonal antibodies (MAbs) targeting the E2 protein of transmissible gastroenteritis (TGE) virus were developed. These MAbs neutralize TGE virus by binding to conserved, conformational epitopes critical for viral infectivity.

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    Area of Science:

    • Virology
    • Immunology
    • Molecular Biology

    Background:

    • Transmissible gastroenteritis (TGE) virus causes significant economic losses in the swine industry.
    • Understanding TGE virus structural proteins and their antigenic properties is crucial for vaccine development and diagnostics.
    • Monoclonal antibodies (MAbs) are valuable tools for characterizing viral proteins and neutralizing viral infectivity.

    Purpose of the Study:

    • To characterize the structural proteins of purified TGE virus.
    • To develop and characterize MAbs specific for TGE virus structural proteins, particularly those involved in neutralization.
    • To identify and map critical neutralizing epitopes on TGE virus and assess their conservation.

    Main Methods:

    • Purification of TGE virus and its structural proteins.

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  • Development of 48 MAbs against TGE virus and characterization of their specificities (E2, N, E1 proteins).
  • Affinity chromatography using MAbs for protein purification.
  • Neutralization assays to determine the efficacy of MAbs.
  • Analysis of MAb-resistant (mar) mutants to identify critical epitopes.
  • Epitope mapping across diverse TGE virus isolates.

    • Main Results:

    • TGE virus comprises three major structural proteins (200,000, 48,000, and 28,000 Da).
    • A panel of 48 MAbs was generated, with 26 specific for the E2 protein, 10 for N, and 3 for E1.
    • Fourteen neutralizing MAbs were specific for the E2 protein, and one MAb (1B.C11) showed neutralization with unknown specificity.
    • Six critical neutralizing epitopes were identified, all conformational, with one (defined by MAb 1G.A7) showing partial resistance to SDS.
    • The frequency of MAb-resistant mutants varied, with most critical epitopes being highly conserved across 11 different TGE virus isolates.

    Conclusions:

    • The E2 protein is a major target for neutralizing antibodies against TGE virus.
    • Highly conserved, conformational epitopes on the E2 protein are critical for TGE virus neutralization.
    • These conserved epitopes represent promising targets for the development of effective TGE vaccines and diagnostic tools.