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

Affinity screening for weak monoclonal antibodies

L Leickt1, A Grubb, S Ohlson

  • 1Department of Natural Sciences, University of Kalmar, Sweden.

Journal of Immunological Methods
|December 5, 1998
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

Permanent Vegetative State: Addendum to a Review by a Working Group Convened by the Royal College of Physicians and Endorsed by the Conference of Medical Royal Colleges.

Journal of the Royal College of Physicians of London·2019
Same author

The identification of discontinuous epitope in the human cystatin C - Monoclonal antibody HCC3 complex.

Journal of proteomics·2018
Same author

Cystatin C: More than a renal function marker.

Revista clinica espanola·2015
Same author

Immobilized cells in microbial nitrate reduction.

Applied biochemistry and biotechnology·2013
Same author

Exploiting weak affinities.

Methods in molecular biology (Clifton, N.J.)·2011
Same author

Serum amyloid a protein and C-reactive protein in normal pregnancy and preeclampsia.

Gynecologic and obstetric investigation·2009

Affinity chromatography effectively screens for low-affinity monoclonal antibodies (mAbs) that are missed by standard assays. This method uses immobilized antigens to select hybridomas producing mAbs with specific low-affinity characteristics.

Area of Science:

  • Biotechnology
  • Immunology
  • Analytical Chemistry

Background:

  • Selecting monoclonal antibodies (mAbs) with specific affinities is crucial for various applications.
  • Traditional immunoassays struggle to identify low-affinity mAbs (dissociation constant (Kd) > 10(-4) M).

Purpose of the Study:

  • To evaluate affinity chromatography as a method for selecting low-affinity monoclonal antibodies.
  • To demonstrate the utility of immobilized antigens in high-performance liquid affinity chromatography (HPLAC) for mAb screening.

Main Methods:

  • High-performance liquid affinity chromatography (HPLAC) was employed.
  • Columns were immobilized with oligosaccharides, dinitrophenol, and digoxin as antigens.
  • Crude monoclonal antibodies in ascites or cell culture supernatants were analyzed.

Related Experiment Videos

Main Results:

  • Monoclonal antibodies showed varying degrees of retardation or adsorption based on their affinity/avidity to immobilized antigens.
  • Antibody retention on the columns correlated with antibody affinity.
  • This allowed for the selection of hybridomas producing desired low-affinity mAbs.

Conclusions:

  • Affinity chromatography is a viable method for selecting low-affinity monoclonal antibodies.
  • HPLAC with immobilized antigens provides a robust approach for screening and selecting hybridomas based on antibody affinity characteristics.