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

Partial purification of human interferon by affinity chromatography.

C B Anfinsen, S Bose, L Corley

    Proceedings of the National Academy of Sciences of the United States of America
    |August 1, 1974
    PubMed
    Summary

    This study demonstrates affinity chromatography for purifying human interferons. Using antibodies against leukocyte interferon effectively purified both leukocyte and fibroblast types, with fibroblast interferon showing higher purity.

    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

    Serendipity in Pediatrics.

    Irish medical journal·2023
    Same author

    Search for dijet resonances in 7 TeV pp collisions at CMS.

    Physical review letters·2011
    Same author

    Charged-particle multiplicity density at midrapidity in central Pb-Pb collisions at sqrt[S(NN)] = 2.76 TeV.

    Physical review letters·2011
    Same author

    Elliptic flow of charged particles in Pb-Pb collisions at sqrt[S(NN)] = 2.76 TeV.

    Physical review letters·2011
    Same author

    Search for quark compositeness with the dijet centrality ratio in pp collisions at √s=7 TeV.

    Physical review letters·2011
    Same author

    Search for stopped Gluinos in pp collisions at square root s=7 TeV.

    Physical review letters·2011

    Area of Science:

    • Biochemistry
    • Immunology
    • Protein Purification

    Background:

    • Human interferons are crucial antiviral proteins produced by various cells.
    • Purification of interferons is essential for studying their properties and therapeutic potential.
    • Existing purification methods often struggle with efficiency and purity.

    Purpose of the Study:

    • To investigate the efficacy of affinity chromatography for purifying human leukocyte and fibroblast interferons.
    • To develop a more efficient method for obtaining highly pure interferon preparations.
    • To compare the purification outcomes for different interferon types.

    Main Methods:

    • Affinity chromatography using Sepharose-4B columns coupled with antibodies to leukocyte interferon.
    • Preparation of antibodies in sheep using partially purified leukocyte interferon as antigen.

    Related Experiment Videos

  • Gel filtration (Sephadex G-100) for initial antigen purification.
  • Addition of carrier proteins (bovine serum albumin, cytochrome c) to eluting buffers for fibroblast interferon purification.

    • Main Results:

    • Both leukocyte and fibroblast interferons were successfully purified from crude tissue culture fluids using the antibody columns.
    • Fibroblast interferon achieved higher purity due to the "common denominator" approach, as fewer fibroblast impurities were recognized by the antibodies.
    • Carrier proteins were necessary for efficient recovery of fibroblast interferon, mitigating adsorption losses.
    • Both interferon types exhibited molecular weights of approximately 20,000-25,000, with some association with higher molecular weight proteins.

    Conclusions:

    • Affinity chromatography with leukocyte interferon antibodies is an effective method for purifying both leukocyte and fibroblast interferons.
    • The "common denominator" approach enhances purification specificity, particularly for fibroblast interferon.
    • Carrier proteins are vital for optimizing the recovery of fibroblast interferon during purification.
    • The study provides a refined method for obtaining pure interferon for further research.