Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Cationic antigens. Problems associated with measurement by ELISA.

A J Pesce, R Apple, N Sawtell

    Journal of Immunological Methods
    |February 27, 1986
    PubMed
    Summary

    Measuring mouse antibody responses to cationized bovine serum albumin (cat BSA) and bovine gammaglobulin (cat BGG) is challenging. Polyanions like heparin effectively reduce non-specific binding and enhance specific antibody detection in immunoassays.

    Related Concept Videos

    You might also read

    Related Articles

    Articles linked to this work by shared authors, journal, and citation graph.

    Sort by
    Same author

    Body mass index and health-related quality of life.

    Obesity science & practice·2018
    Same author

    A study of genotyping for management of human papillomavirus-positive, cytology-negative cervical screening results.

    Journal of clinical microbiology·2014
    Same author

    Microencapsulation: the future of oral immunotherapy?

    BioDrugs : clinical immunotherapeutics, biopharmaceuticals and gene therapy·2007
    Same author

    Factor V Leiden polymorphism and the rate of fibrosis development in chronic hepatitis C virus infection.

    Gut·2003
    Same author

    Effect of outliers and nonhealthy individuals on reference interval estimation.

    Clinical chemistry·2001
    Same author

    Amiodarone in the treatment of junctional ectopic tachycardia after cardiac surgery in children: report of two cases and review of the literature.

    American journal of therapeutics·2001

    Area of Science:

    • Immunology
    • Biochemistry
    • Assay Development

    Background:

    • Measuring mouse antibody responses to cationized proteins presents unique challenges due to antigen properties.
    • Cationized bovine gammaglobulin (cat BGG) exhibited non-specific binding with enzyme-conjugated antibodies.
    • Cationized bovine serum albumin (cat BSA) showed non-specific reactions, though specific antibody reactions could be enhanced.

    Purpose of the Study:

    • To investigate and overcome complications in measuring mouse antibody responses to cationized antigens.
    • To identify methods for minimizing non-specific binding and enhancing specific antibody detection in immunoassays.
    • To propose a model explaining the observed interactions and guide future assay development.

    Main Methods:

    • Enzyme-linked immunosorbent assays (ELISA) were employed to measure antibody responses.

    Related Experiment Videos

  • The effects of polyanions (heparin, dextran sulfate) on antigen-antibody reactions were evaluated.
  • ELISA inhibition experiments were conducted using varying antigen concentrations.

    • Main Results:

    • Heparin addition minimized non-specific binding of rabbit anti-mouse gammaglobulin to cat BGG.
    • Heparin and dextran sulfate enhanced specific antibody reactions with cat BSA while reducing non-specific binding.
    • High antigen concentrations (>100 µg/ml) in ELISA inhibition tests led to non-specific inhibition.

    Conclusions:

    • A polycationic surface model explains the observed non-specific and specific binding phenomena.
    • Polyanions neutralize the cationic surface, reducing non-specific reactions and enhancing specific antibody detection.
    • Consideration of polycationic molecule properties is crucial for accurate antibody measurement in immunoassays.