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Titolo:
Construction and functional evaluation of a single-chain antibody fragmentthat neutralizes toxin Aahl from the venom of the scorpion Androctonus australis hector
Autore:
Devaux, C; Moreau, E; Goyffon, M; Rochat, H; Billiald, P;
Indirizzi:
Univ Mediterranee, Fac Med Nord, IFR Jean Roche, CNRS,UMR 6560, F-13916 Marseille 20, France Univ Mediterranee Marseille France 20 6560, F-13916 Marseille 20, France Museum Natl Hist Nat, F-75231 Paris, France Museum Natl Hist Nat Paris France F-75231 ist Nat, F-75231 Paris, France Fac Pharm, Tours, France Fac Pharm Tours FranceFac Pharm, Tours, France
Titolo Testata:
EUROPEAN JOURNAL OF BIOCHEMISTRY
fascicolo: 3, volume: 268, anno: 2001,
pagine: 694 - 702
SICI:
0014-2956(200102)268:3<694:CAFEOA>2.0.ZU;2-0
Fonte:
ISI
Lingua:
ENG
Soggetto:
MONOCLONAL-ANTIBODIES; ESCHERICHIA-COLI; BINDING; SCFV; FV; PURIFICATION; NEUROTOXINS; HEMOCYANIN; AFFINITY; LIBRARY;
Keywords:
neurotoxins; neutralization; scFv; scorpion venom;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
42
Recensione:
Indirizzi per estratti:
Indirizzo: Devaux, C Univ Mediterranee, Fac Med Nord, IFR Jean Roche, CNRS,UMR 6560, Blvd P Dramard, F-13916 Marseille 20, France Univ Mediterranee Blvd P Dramard Marseille France 20 20, France
Citazione:
C. Devaux et al., "Construction and functional evaluation of a single-chain antibody fragmentthat neutralizes toxin Aahl from the venom of the scorpion Androctonus australis hector", EUR J BIOCH, 268(3), 2001, pp. 694-702

Abstract

9C2 is a murine monoclonal IgG that participates in the neutralization of Androctonus australis hector scorpion venom. It recognizes AahI and AahIII,two of the three main neurotoxins responsible for almost all the toxicity of the venom when injected into mammals. Using PCR we cloned the antibody variable region coding genes from 9C2 hybridoma cells and constructed a geneencoding a single chain antibody variable fragment molecule (scFv). This scFv was produced in the periplasm of Escherichia coli in a soluble and functional form and purified in a single step using protein L-agarose beads yielding 1-2 mg.L-1 of bacterial culture. scFv9C2 was predominantly monomeric but also tended to form dimeric and oligomeric structures, all capable of binding toxin AahI. The affinity of scFv and the parental mAb for toxin AahIand homologous toxin AahIII was of the same magnitude, in the nanomolar range. Similarly, purified forms of scFv9C2 completely inhibited the binding of toxin AahI to rat brain synaptosomes. Finally, scFv9C2 was efficient in protecting mice against the toxic effects of AahI after injection of the toxin and scFv to mice by the intracerebroventricular route in a molar ratio as low as 0.36 : 1. Thus, we produced a recombinant scFv that reproduces the recognition properties of the parent antibody and neutralizes the scorpion neurotoxin AahI, thereby opening new prospects for the treatment of envenomation.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/11/20 alle ore 13:44:38