Anti-idiotypic antibody to an anti-alprenolol monoclonal antibody: promotion of ligand binding to the idiotype
Abstract
The authors previously described the production of four monoclonal antibodies to the beta-adrenergic receptor antagonist alprenolol. One of these antibodies, 5B7 (IgG/sub 2a/k), was used to generate polyclonal anti-idiotypic antisera in rabbits. In contrast to the predicted results, the anti-idiotypic antisera (R-9) promoted (/sup 125/I)-CYP binding to the monoclonal antibody 5B7. In the presence of R-9 antisera (1:300 dilution), the binding affinity improved with the dissociation constant decreasing 100-fold from 10nM to 0.1nM. This effect could not be reproduced with pre-immune, rabbit anti-mouse, or antiidiotypic antisera generated to monoclonal antibodies of different specificity. Furthermore, R-9 alone did not bind ligand. The R-9 IgG fraction was isolated by DEAE-cellulose chromatography then adsorbed to a 5B7-immunoaffinity resin and eluted with 0.5 M NaCl. This yielded a fraction with enhanced binding activity. F(ab) fragments of 5B7 and R-9 produced the same effect indicating that polyvalency was not necessary for the enhanced ligand binding. Finally, the ligand alprenolol promoted the binding of (/sup 125/I)-5B7 to %-9 as assessed by fractionating the resulting soluble complex by size exclusion chromatography on a TSK-3000 HPLC column. These results demonstrate that anti-idiotypic antibodies can be generated which will promote the binding of antigen to the original idiotype.
- Authors:
- Publication Date:
- Research Org.:
- Harvard Medical School, Boston, MA
- OSTI Identifier:
- 5278299
- Report Number(s):
- CONF-8604222-
Journal ID: CODEN: FEPRA; TRN: 86-030448
- Resource Type:
- Conference
- Journal Name:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
- Additional Journal Information:
- Journal Volume: 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; MONOCLONAL ANTIBODIES; BIOCHEMICAL REACTION KINETICS; AFFINITY; CHROMATOGRAPHY; IODINE 125; LIGANDS; RABBITS; TRACER TECHNIQUES; ANIMALS; ANTIBODIES; BETA DECAY RADIOISOTOPES; DAYS LIVING RADIOISOTOPES; ELECTRON CAPTURE RADIOISOTOPES; INTERMEDIATE MASS NUCLEI; IODINE ISOTOPES; ISOTOPE APPLICATIONS; ISOTOPES; KINETICS; MAMMALS; NUCLEI; ODD-EVEN NUCLEI; RADIOISOTOPES; REACTION KINETICS; SEPARATION PROCESSES; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques
Citation Formats
Sawutz, G, and Homcy, C J. Anti-idiotypic antibody to an anti-alprenolol monoclonal antibody: promotion of ligand binding to the idiotype. United States: N. p., 1986.
Web.
Sawutz, G, & Homcy, C J. Anti-idiotypic antibody to an anti-alprenolol monoclonal antibody: promotion of ligand binding to the idiotype. United States.
Sawutz, G, and Homcy, C J. 1986.
"Anti-idiotypic antibody to an anti-alprenolol monoclonal antibody: promotion of ligand binding to the idiotype". United States.
@article{osti_5278299,
title = {Anti-idiotypic antibody to an anti-alprenolol monoclonal antibody: promotion of ligand binding to the idiotype},
author = {Sawutz, G and Homcy, C J},
abstractNote = {The authors previously described the production of four monoclonal antibodies to the beta-adrenergic receptor antagonist alprenolol. One of these antibodies, 5B7 (IgG/sub 2a/k), was used to generate polyclonal anti-idiotypic antisera in rabbits. In contrast to the predicted results, the anti-idiotypic antisera (R-9) promoted (/sup 125/I)-CYP binding to the monoclonal antibody 5B7. In the presence of R-9 antisera (1:300 dilution), the binding affinity improved with the dissociation constant decreasing 100-fold from 10nM to 0.1nM. This effect could not be reproduced with pre-immune, rabbit anti-mouse, or antiidiotypic antisera generated to monoclonal antibodies of different specificity. Furthermore, R-9 alone did not bind ligand. The R-9 IgG fraction was isolated by DEAE-cellulose chromatography then adsorbed to a 5B7-immunoaffinity resin and eluted with 0.5 M NaCl. This yielded a fraction with enhanced binding activity. F(ab) fragments of 5B7 and R-9 produced the same effect indicating that polyvalency was not necessary for the enhanced ligand binding. Finally, the ligand alprenolol promoted the binding of (/sup 125/I)-5B7 to %-9 as assessed by fractionating the resulting soluble complex by size exclusion chromatography on a TSK-3000 HPLC column. These results demonstrate that anti-idiotypic antibodies can be generated which will promote the binding of antigen to the original idiotype.},
doi = {},
url = {https://www.osti.gov/biblio/5278299},
journal = {Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)},
number = ,
volume = 45:3,
place = {United States},
year = {Sat Mar 01 00:00:00 EST 1986},
month = {Sat Mar 01 00:00:00 EST 1986}
}