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Titolo:
Functional characterization of two anti-estradiol antibodies as deduced from modelling and site-directed mutagenesis experiments
Autore:
Bettsworth, F; Monnet, C; Watelet, B; Battail-Poirot, N; Gilquin, B; Jolivet, M; Menez, A; Arnaud, M; Ducancel, F;
Indirizzi:
CEA Saclay, Dept Ingn & Etud Prot, F-91191 Gif Sur Yvette, France CEA Saclay Gif Sur Yvette France F-91191 F-91191 Gif Sur Yvette, France bioMerieux, Dept R&D Immunoessais, F-69280 Marcy Etoile, France bioMerieux Marcy Etoile France F-69280 ais, F-69280 Marcy Etoile, France
Titolo Testata:
JOURNAL OF MOLECULAR RECOGNITION
fascicolo: 2, volume: 14, anno: 2001,
pagine: 99 - 109
SICI:
0952-3499(200103/04)14:2<99:FCOTAA>2.0.ZU;2-Z
Fonte:
ISI
Lingua:
ENG
Soggetto:
SINGLE-CHAIN FV; COMPLEMENTARITY-DETERMINING REGIONS; RECEPTOR-MIMICKING ANTIBODY; 3-DIMENSIONAL STRUCTURE; MONOCLONAL-ANTIBODY; ESCHERICHIA-COLI; PHAGE DISPLAY; HIGH-AFFINITY; BINDING SITE; HYPERVARIABLE LOOPS;
Keywords:
antibody; anti-hapten; estradiol; scFv; modelling; site-directed mutagenesis;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
53
Recensione:
Indirizzi per estratti:
Indirizzo: Ducancel, F CEA Saclay, Dept Ingn & Etud Prot, F-91191 Gif Sur Yvette, France CEA Saclay Gif Sur Yvette France F-91191 Sur Yvette, France
Citazione:
F. Bettsworth et al., "Functional characterization of two anti-estradiol antibodies as deduced from modelling and site-directed mutagenesis experiments", J MOL RECOG, 14(2), 2001, pp. 99-109

Abstract

Monoclonal antibodies are now widely used to measure the concentration of steroid hormones in human serum samples. The great development of molecularengineering techniques over the past 10 years has made possible the improvement of specificity and/or sensitivity of selected antibodies. We have obtained two monoclonal antibodies, 17E12E5 and 10G6D6, using estradiol-6-ethyl methoxy carbonyl (EMC)-bovine serum albumin (BSA) as immunogen, To tentatively improve their affinities for natural estradiol, we have initiated their structural and functional studies. For this purpose, we have cloned and sequenced the genes encoding the variable fragments of each antibody. Single chain variable fragments (scFv) were produced into the periplasmic space of E. coli using the pLIP6 expression vector. Mapping of the functional structures of both antibodies was obtained by combination of modelling and mutational analyses together with crossreaction studies. The two binding pockets are described and models of estradiol complexed to 17E12E5 and 10G6D6 are proposed. Copyright (C) 2001 John Wiley & Sons, Ltd.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 22/01/20 alle ore 18:44:01