Accession Number:

ADA180670

Title:

Use of Fluorodinitrobenzene to Identify Monoclonal Antibodies Which are Suitable for Conjugation to Periodate-Oxidized Horseradish Peroxidase,

Descriptive Note:

Corporate Author:

WALTER REED ARMY INST OF RESEARCH WASHINGTON DC

Personal Author(s):

Report Date:

1985-01-01

Pagination or Media Count:

7.0

Abstract:

During the recent development of a double antibody enzyme-linked immunosorbent assay for detecting malaria sporozoites in mosquitoes it was found that a large number of potentially useful monoclonal antibodies lost their capacity for binding antigen after conjugation to periodate-oxidized horseradish peroxidase HPO. Since HPO reacts with primary amino groups, we used a simple chemical reaction with fluorodinitrobenzene FDNB to determine if the loss of antigen binding was due to the requirement for unmodified primary amino groups in the binding site. FDNB-treated antibodies which reacted with antigen in an indirect fluorescent antibody assay IFA also yielded successful HPO-antibody conjugates. Conversely, those antibodies which did not react with antigen after treatment with FDNB failed to produce useful HPO-antibody conjugates. These data suggest that conjugation of oxidized HPO to primary amines in or near the antigen-combining site yields conjugates in which the antibody is inactive. Use of FDNB to predict the suitability of antibodies for conjugation to periodate-oxidized HPO may save a considerable amount of time over randomly selecting antibodies for conjugation and testing. Reprints

Subject Categories:

  • Biochemistry
  • Medicine and Medical Research

Distribution Statement:

APPROVED FOR PUBLIC RELEASE