PMID: 2507642Sep 29, 1989Paper

Use of bispecific hybrid antibodies for the development of a homogeneous enzyme immunoassay

Journal of Immunological Methods
G GörögF Celada

Abstract

Hybrid bispecific monoclonal antibodies reacting with carcinoembryonal antigen (CEA) and with the E. coli enzyme beta-galactosidase (GZ) were produced by fusion of hybridomas or chemical linkage of half-antibodies. Since the original anti-GZ antibody used in these experiments was capable of protecting GZ from thermal denaturation, it was possible, by hybridizing it with two different non-competitive anti-CEA antibodies, to design a homogeneous enzyme immunoassay for quantitation of CEA. In fact, a mathematical analysis of the reaction indicates that, under appropriate concentrations of the reactants, circular complexes can be formed which contain the two hybrid antibodies, the GZ enzyme and the CEA antigen. The stability of these complexes can be expected to be substantially greater than that of the more labile CEA-free GZ-antibody complexes, prompting a significant increase in the amount of enzyme molecules which are bound to antibody and are consequently protected from thermal denaturation. These expectations were supported by experimental results: under appropriate conditions, heat-resistant enzyme activity was indeed proportional to concentration of CEA in the range up to 75 ng/ml. As predicted by theory, however, in the pr...Continue Reading

References

Aug 5, 1992·Journal of Molecular Biology·L JinJ A Wells
Oct 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·M R SureshC Milstein
Jan 1, 1987·European Journal of Immunology·A Lanzavecchia, D Scheidegger
May 26, 1972·Biochemical and Biophysical Research Communications·K E RubensteinE F Ullman
Oct 6, 1983·Nature·C Milstein, A C Cuello
Feb 1, 1980·Analytical Biochemistry·I GibbonsE F Ullman
Apr 1, 1981·Proceedings of the National Academy of Sciences of the United States of America·R S AccollaF Celada

❮ Previous
Next ❯

Citations

Jan 1, 1992·International Journal of Clinical & Laboratory Research·O Nolan, R O'Kennedy
Jul 6, 1992·Journal of Immunological Methods·S A FernandoG S Wilson
May 16, 1994·Journal of Immunological Methods·A G Cook, P J Wood
Dec 15, 1995·Journal of Immunological Methods·B S DeSilva, G S Wilson
Oct 1, 1992·Molecular Immunology·W J AllardM T Largen
Feb 1, 1991·Current Opinion in Biotechnology·T T Ngo
Oct 5, 2001·The Journals of Gerontology. Series A, Biological Sciences and Medical Sciences·J VettrainoR Arking
Oct 6, 2000·Methods : a Companion to Methods in Enzymology·B S DeSilva, G S Wilson

❮ Previous
Next ❯

Related Concepts

Related Feeds

Antibody Specificity

Antibodies produced by B cells are highly specific for antigen as a result of random gene recombination and somatic hypermutation and affinity maturation. As the main effector of the humoral immune system, antibodies can neutralize foreign cells. Find the latest research on antibody specificity here.