Production of scFv recombinant fragments against 2,4-dichlorophenoxyacetis acid hapten using naive phage library

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F03%3A00008470" target="_blank" >RIV/00027162:_____/03:00008470 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Production of scFv recombinant fragments against 2,4-dichlorophenoxyacetis acid hapten using naive phage library

Popis výsledku v původním jazyce

Three single chain variable fragment (scFv) antibodies against 2,4-dichlophenoxyacetic acid (2,4-D) herbicide were produced by the Griffin1.library. The selection of the scFv from the phage library was carried out by 2,4-D-protein coated tubes with different levels of hapten substitution in the conjugate. The scFv phage clones were isolated within the five round library panning and the antibodies were expressed in Escherichia coli HB2151. The recombinant products were purified by metal affinity chromatography yielding 200 ?g of pure scFv per 1 liter of bacterial culture. The antibody fragments provided steep curves in conventional indirect ELISA having the IC50 values from 10.2 to 14.5 ng/ml established for 2,4-D standard. Interestingly enough, the recombinant ScFv E1 anti-body exhibited 68% cross-reactivity with 2,4-dichlorphenol (2,4-D = 100%), and 38.0% with methylchlorophenoxy-acetic acid (MCPA) whereas reaction with other phenoxyacetic compounds was low. Similar characteristics we

Název v anglickém jazyce

Production of scFv recombinant fragments against 2,4-dichlorophenoxyacetis acid hapten using naive phage library

Popis výsledku anglicky

Three single chain variable fragment (scFv) antibodies against 2,4-dichlophenoxyacetic acid (2,4-D) herbicide were produced by the Griffin1.library. The selection of the scFv from the phage library was carried out by 2,4-D-protein coated tubes with different levels of hapten substitution in the conjugate. The scFv phage clones were isolated within the five round library panning and the antibodies were expressed in Escherichia coli HB2151. The recombinant products were purified by metal affinity chromatography yielding 200 ?g of pure scFv per 1 liter of bacterial culture. The antibody fragments provided steep curves in conventional indirect ELISA having the IC50 values from 10.2 to 14.5 ng/ml established for 2,4-D standard. Interestingly enough, the recombinant ScFv E1 anti-body exhibited 68% cross-reactivity with 2,4-dichlorphenol (2,4-D = 100%), and 38.0% with methylchlorophenoxy-acetic acid (MCPA) whereas reaction with other phenoxyacetic compounds was low. Similar characteristics we

Druh

J_x - Nezařazeno - Článek v odborném periodiku (Jimp, Jsc a Jost)

CEP obor

EI - Biotechnologie a bionika

OECD FORD obor

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Projekt

<a href="/cs/project/GA525%2F00%2F1261" target="_blank" >GA525/00/1261: Produkce rekombinantních protilátek pro analýzu životního prostředí a potravin</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2003

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Veterinární medicína

ISSN

0375-8427

e-ISSN

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Svazek periodika

48

Číslo periodika v rámci svazku

9

Stát vydavatele periodika

CZ - Česká republika

Počet stran výsledku

11

Strana od-do

237-247

Kód UT WoS článku

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EID výsledku v databázi Scopus

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