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Related Experiment Videos

Mapping of viral conformational epitopes using biosensor measurements

H Saunal1, M H Van Regenmortel

  • 1UPR 9021 Immunochimie des Peptides et des Virus, Institut de Biologie Moléculaire and Cellulaire, CNRS, Strasbourg, France.

Journal of Immunological Methods
|June 14, 1995
PubMed
Summary

Biosensor technology reveals new locations for tobacco mosaic virus epitopes. This advanced method maps conformational epitopes more effectively than traditional immunoassays, identifying neotopes and metatopes on viral surfaces.

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Area of Science:

  • Virology
  • Structural Biology
  • Immunology

Background:

  • Previous electron microscopy suggested neotopes (quaternary structure-specific) were along the entire virus, while metatopes (present in intact and dissociated forms) were at one extremity.
  • Classical immunoassays had limitations in precisely mapping conformational epitopes on viral surfaces.

Purpose of the Study:

  • To investigate the binding properties of antibodies to neotopes and metatopes on tobacco mosaic virus using BIAcore biosensor technology.
  • To identify novel locations of neotopes and metatopes on the viral surface.
  • To explore the induction of neotope specificity in dissociated viral subunits.

Main Methods:

  • Utilized BIAcore biosensor technology for real-time analysis of antibody-antigen interactions.

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  • Performed capture assays with viral subunits to determine epitope localization.
  • Conducted binding stoichiometry calculations to quantify epitope presence.
  • Employed two-site binding assays to investigate epitope induction.

    • Main Results:

    • Demonstrated the presence of neotope and metatope specificities on additional viral surface areas not previously identified.
    • Quantified epitope distribution through binding stoichiometry and capture assays.
    • Showed that neotope specificity can be induced in dissociated viral subunits upon binding of an antimetatope antibody.

    Conclusions:

    • Biosensor technology offers superior capabilities for mapping conformational epitopes in viral proteins compared to traditional methods.
    • The study refined the understanding of epitope distribution on the tobacco mosaic virus particle.
    • Revealed dynamic epitope expression and potential for conformational changes in viral subunits.