Measurement of affinity of viral monoclonal antibodies using Fab'-peroxidase conjugate. Influence of antibody concentration on apparent affinity.
The binding affinity of a monoclonal antibody to tobacco mosaic virus (TMV) was studied using a Fab'-peroxidase conjugate. Measurement of the enzymatic activity allowed the determination of the amount of free antibody present after ultracentrifugation of virus-antibody complexes at equilibrium. The method was very sensitive and allowed measurements over a 1000-fold range of antibody concns. The calculated affinity constant decreased about 25 fold when the antibody concn used in the binding assay was increased from 30 ng/ml to 35 micrograms/ml.
Azimzadeh
A
A
Laboratoire d'Immunochimie, CNRS, Strasbourg, France.
Weiss
E
E
Van Regenmortel
M H
MH
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D007140:Immunoglobulin Fab Fragments / Q000276:immunology*; D066298:In Vitro Techniques; D010544:Peroxidases / Q000494:pharmacology*; D014027:Tobacco Mosaic Virus / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D010544:Peroxidases