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Titolo:
Immunolabeling of CD3-positive lymphocytes with a recombinant single-chainantibody/alkaline phosphatase conjugate
Autore:
Bourin, P; Servat, A; Lataillade, JJ; Goyffon, M; Vaux, D; Billiald, P;
Indirizzi:
Museum Natl Hist Nat, LERAI, F-75231 Paris 05, France Museum Natl Hist Nat Paris France 05 at, LERAI, F-75231 Paris 05, France Ctr Transfus Sanguine Armees, F-92140 Clamart, France Ctr Transfus Sanguine Armees Clamart France F-92140 2140 Clamart, France Fac Pharm, F-37200 Tours, France Fac Pharm Tours France F-37200Fac Pharm, F-37200 Tours, France Univ Oxford, Sir William Dunn Sch Pathol, Oxford OX1 3RE, England Univ Oxford Oxford England OX1 3RE n Sch Pathol, Oxford OX1 3RE, England
Titolo Testata:
BIOLOGICAL CHEMISTRY
fascicolo: 2, volume: 381, anno: 2000,
pagine: 173 - 178
SICI:
1431-6730(200002)381:2<173:IOCLWA>2.0.ZU;2-Q
Fonte:
ISI
Lingua:
ENG
Soggetto:
TRUNCATED DIPHTHERIA-TOXIN; T-CELL; ESCHERICHIA-COLI; FUSION-PROTEINS; IN-VITRO; ANTIBODIES; CD3; ANTIGEN; FV; CONSTRUCTION;
Keywords:
bacterial expression; immunoconjugate; recombinant antibody; scFv;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
27
Recensione:
Indirizzi per estratti:
Indirizzo: Billiald, P Museum Natl Hist Nat, LERAI, 57 Rue Cuvier, F-75231 Paris 05, France Museum Natl Hist Nat 57 Rue Cuvier Paris France 05 05, France
Citazione:
P. Bourin et al., "Immunolabeling of CD3-positive lymphocytes with a recombinant single-chainantibody/alkaline phosphatase conjugate", BIOL CHEM, 381(2), 2000, pp. 173-178

Abstract

G3(3) is a novel murine monoclonal antibody directed against the CD3 antigen of human T lymphocytes which could be used to analyze lymphoid malignancies. We have produced and characterized a recombinant colorimetric immunoconjugate with the antigen-binding specificity of antibody G3(3). A gene encoding a single-chain antibody variable fragment (scFv) was assembled using the original hybridoma cells as a source of antibody variable heavy OIH) andvariable light (VL) chain genes. The chimeric gene was introduced into a prokaryotic expression vector in order to produce a soluble scFv fused to bacterial alkaline phosphatase, DNA sequencing and Western blotting analyses demonstrated the integrity of the soluble immunoconjugate recovered from induced recombinant bacteria. The scFv/AP protein was bifunctional and similar in immunoreactivity to the parent G3(3) antibody. Flow cytometry and immunostaining experiments confirmed that the activity of the scFv/AP protein compares favourably with that of the parent antibody. The scFv/AP conjugate was bound to CD3 antigen at the surface of T cells and was directly detected by its enzymatic activity. Thus this novel fusion protein has potential applications as an immunodiagnostic reagent.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/11/20 alle ore 13:59:33