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

A solid-phase immunofluorescence assay (SIFA) for screening antigen-specific hybridomas.

B Micheel, U Karsten, H Fiebach

    Journal of Immunological Methods
    |January 1, 1981
    PubMed
    Summary

    A novel solid-phase immunofluorescence assay (SIFA) efficiently screens monoclonal hybridoma antibodies. This method requires minimal antigen and processes numerous samples rapidly, offering comparable sensitivity to radioimmunoassays.

    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

    Ecophysiological study of green microalgae isolated from the grit crust of the Atacama Desert.

    Journal of phycology·2026
    Same author

    Multi-omics for studying and understanding polar life.

    Nature communications·2023
    Same author

    Erratum to "Cluster binding studies with two anti-Thomsen-Friedenreich (anti-core-1, CD176) antibodies: Evidence for a quadruple epitope" [Int. Immunopharmacol. 72 (2019) 186-194].

    International immunopharmacology·2021
    Same author

    Cluster binding studies with two anti-Thomsen-Friedenreich (anti-core-1, CD176, TF) antibodies: Evidence for a multiple TF epitope.

    International immunopharmacology·2019
    Same author

    Photosynthetic performance of Arctic macroalgae after transplantation from deep to shallow waters.

    Oecologia·2017
    Same author

    Ultraviolet-absorbing scytonemin and mycosporine-like amino acid derivatives in exposed, rock-inhabiting cyanobacterial lichens.

    Oecologia·2017

    Area of Science:

    • Immunology
    • Biochemistry

    Background:

    • Hybridoma technology is crucial for producing monoclonal antibodies.
    • Screening hybridoma supernatants for specific antibody production is a critical step.
    • Existing screening methods can be time-consuming or require substantial reagents.

    Purpose of the Study:

    • To describe a novel solid-phase immunofluorescence assay (SIFA) for screening monoclonal hybridoma antibodies.
    • To evaluate the efficiency, material requirements, and throughput of the SIFA.
    • To compare the sensitivity of SIFA with established radioimmunoassay methods.

    Main Methods:

    • Development of a solid-phase immunofluorescence assay using microtitration plates and cellulose nitrate discs.
    • Incubation steps involved anti-mouse IgG, hybridoma samples, and FITC-labeled soluble antigen.

    Related Experiment Videos

  • Quantification of fluorescence intensity using a microfluorometer.
  • Application of the assay for selecting hybridomas against human serum albumin.

    • Main Results:

    • The SIFA is simple and requires minimal labeled antigen (20-100 ng/sample).
    • The assay allows for high throughput, testing 500-1000 samples within 2 days.
    • The SIFA demonstrated sensitivity comparable to radioimmunoassay when tested with hybridomas against human serum albumin.

    Conclusions:

    • Solid-phase immunofluorescence assay (SIFA) provides an efficient and sensitive method for screening monoclonal hybridoma antibodies against soluble antigens.
    • The assay's low material requirement and high throughput make it suitable for large-scale hybridoma selection.
    • SIFA offers a viable alternative to traditional radioimmunoassays for antibody screening.