Related Experiment Video

Updated: May 19, 2026

Generation of Monoclonal Antibodies Against Natural Products
12:15

Generation of Monoclonal Antibodies Against Natural Products

Published on: April 6, 2019

Establishment of reverse direct ELISA and its application in screening high-affinity monoclonal antibodies

Chunmei Zhang1, Rende Li, Yongming Li

  • 1Department of Immunology, The Fourth Military Medical University, Xi'an, Shaanxi Province, P.R. China.

Hybridoma (2005)
|August 17, 2012
PubMed

Traditionally monoclonal antibody (MAb) titer is determined by indirect enzyme-linked immunosorbent assay (ELISA), which is primarily used to evaluate the quality of MAbs. In this study, the titer and affinity of a group of MAbs against ovalbumin (OVA) were tested by indirect ELISA and the ELISA method reported previously. Data showed that there may be great differences between the indirect ELISA antibody titer and affinity value of MAbs. For the first time, a simple and effective reverse direct ELISA (RD-ELISA) was established for the detection of high-affinity MAbs. Among the group of MAbs to OVA, a certain proportion of antibodies with high affinity but low indirect ELISA titer do exist and can be clearly and efficiently detected by RD-ELISA. This study demonstrates that RD-ELISA is an effective method for high-affinity MAb screening.

More Related Videos

Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations09:07

Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations

Published on: August 22, 2019

Detection of Antibodies That Neutralize the Cellular Uptake of Enzyme Replacement Therapies with a Cell-based Assay07:52

Detection of Antibodies That Neutralize the Cellular Uptake of Enzyme Replacement Therapies with a Cell-based Assay

Published on: September 10, 2018

Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or quantified.
Hybridoma Technology01:31

Hybridoma Technology

Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
Hybridoma Selection
Commonly used fusion techniques — electroporation, polyethylene glycol...