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

Immunization procedures for E. coli proteins.

V R Anicetti1, M A Simonetti, L L Blackwood

  • 1Department of Medicinal and Analytical Chemistry, Genentech, Inc., South San Francisco, CA 94080.

Applied Biochemistry and Biotechnology
|November 1, 1989
PubMed
Summary
This summary is machine-generated.

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

Redox-driven mineral and organic associations in Jezero Crater, Mars.

Nature·2025
Same author

"More Than Just the Two Percent": The reproductive politics of envisioning abortion and miscarriage in Turner syndrome.

Medical anthropology quarterly·2025
Same author

Impact of COVID-19 on diagnosis and testing for TB in a high-resource, low-burden setting.

The international journal of tuberculosis and lung disease : the official journal of the International Union against Tuberculosis and Lung Disease·2022
Same author

Systematic review of the evidence for sustained efficacy of dietary interventions for reducing appetite or energy intake.

Obesity reviews : an official journal of the International Association for the Study of Obesity·2018
Same author

High MMP-1, MMP-2, and MMP-9 protein levels in osteoarthritis.

Genetics and molecular research : GMR·2015
Same author

When pneumonia does not respond to antibiotics: a challenging neonatal diagnosis.

Archives of disease in childhood. Education and practice edition·2014

The cascade immunization method significantly improved antibody production against minor components in complex protein mixtures. This enhanced antibody response was observed after 112 days, highlighting the protocol

Area of Science:

  • Immunology
  • Protein Chemistry
  • Biotechnology

Background:

  • Producing antibodies against minor components in complex protein mixtures, such as E. coli protein (ECP), is challenging due to the dominance of highly immunogenic proteins.
  • Conventional immunization protocols may not effectively elicit responses against less abundant antigens within a mixture.

Purpose of the Study:

  • To compare three immunization procedures for antibody production against minor components of a complex E. coli protein (ECP) mixture.
  • To evaluate the efficacy of selective in vitro (cascade) and in vivo (passive) depletion methods compared to a conventional protocol.

Main Methods:

  • Comparison of a conventional immunization protocol with cascade (in vitro depletion) and passive (in vivo depletion) methods for producing antibodies to ECP.

Related Experiment Videos

  • Indirect ELISA was used to determine antibody titers at 60 days post-immunization.
  • Two-dimensional SDS-PAGE immunoblots were employed to analyze antibody reactivity to minor ECP components at 112 days.

    • Main Results:

    • Maximum ELISA antibody titers were achieved at 60 days across all protocols.
    • Significant antibody reactivity to minor ECP components was only detected at 112 days.
    • The cascade immunization procedure demonstrated a marked improvement in antibody response to minor components compared to other methods.

    Conclusions:

    • The cascade immunization protocol, utilizing at least three hosts and the methods of Thalhamer and Freund, is preferred for generating antibodies against minor components in complex antigen mixtures like ECPs.
    • Characterization of antisera is best achieved using silver-stained two-dimensional SDS-PAGE and immunoblotting for detailed analysis.