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 presence of excess CEA - in fact at CEA concentrations which are higher than those of possible clinical relevance - circular complexes tended to open up, leading to a marked prozone effect.

Use of bispecific hybrid antibodies for the development of a homogeneous enzyme immunoassay / Gorog, G; Gandolfi, A; Paradisi, G; Rolleri, E; Klasen, E; Dessi', V; Strom, Roberto; Celada, F.. - In: JOURNAL OF IMMUNOLOGICAL METHODS. - ISSN 0022-1759. - STAMPA. - 123:1(1989), pp. 131-140. [10.1016/0022-1759(89)90037-9]

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

STROM, Roberto;
1989

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 presence of excess CEA - in fact at CEA concentrations which are higher than those of possible clinical relevance - circular complexes tended to open up, leading to a marked prozone effect.
1989
Carcinoembryonic antigen; Hybrid antibodies; Hybridomas; Monoclonal antibodies; Immunoenzymatic technology
01 Pubblicazione su rivista::01a Articolo in rivista
Use of bispecific hybrid antibodies for the development of a homogeneous enzyme immunoassay / Gorog, G; Gandolfi, A; Paradisi, G; Rolleri, E; Klasen, E; Dessi', V; Strom, Roberto; Celada, F.. - In: JOURNAL OF IMMUNOLOGICAL METHODS. - ISSN 0022-1759. - STAMPA. - 123:1(1989), pp. 131-140. [10.1016/0022-1759(89)90037-9]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11573/414665
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