Generation and characterization of single-chain antibody fragments specific against transmembrane envelope glycoprotein gp46 of Maedi-visna Virus

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216224%3A14310%2F04%3A00010104" target="_blank" >RIV/00216224:14310/04:00010104 - isvavai.cz</a>

Alternative codes found

RIV/62157124:16170/04:00000151

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

Generation and characterization of single-chain antibody fragments specific against transmembrane envelope glycoprotein gp46 of Maedi-visna Virus

Original language description

A single-chain antibody fragments (scFv) was developed directed against transmembrane envelope glycoprotein gp46 of the virus maedi-visna, by the application of the antibody phage display library. To get specific scFv binders, the library was panned against the biotinylated peptide of 20 amino acids corresponding to the principal immunodominant domain of gp46 protein. The number of positively binding scFvs was evaluated by scFv-phage ELISA, BstN1 fingerprinting and DNA sequencing. The scFvs were expressed in soluble form and purified by immobilized metal affinity chromatography (IMAC) with a yield of 2 to 2.5 mg/l. Two scFvs have shown to recognize gp46 and gp150 proteins in Western blot analysis. The scFvs also recognized the virus in infected cells as shown by immunofluorescence assay. The affinity of the obtained antibody fragments to gp46 peptide was measured by surface plasmon resonance, and the resulting KA was in the 10^6 to 10^7 l/mol range. The application of characterized scF

Czech name

Produkce a charakterisace scFv protilátek specifických proti transmembránovému obalovému glykoproteinu viru Maedi-visna

Czech description

Byla připravena scFv protilátka proti transmembránovému glykoproteinu viru Maedi visna použitím genetické fágové knihovny DNA fragmentů. Selekce se prováděla pomocí biotinylovaného peptidu z 20 aminokyselin, který odpovídal imunodominantní části proteinugp46. Protilátky ve formě scFv byly přečištěny metaloafinitní chromatografií a charakterisovány pomocí biosensoru na bázi povrchové resonance plasmonů Spreeta.

Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

CE - Biochemistry

OECD FORD branch

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Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

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

Publication year

2004

Confidentiality

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

Name of the periodical

Journal of Virological Methods

ISSN

0166-0934

e-ISSN

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Volume of the periodical

115

Issue of the periodical within the volume

1

Country of publishing house

CZ - CZECH REPUBLIC

Number of pages

10

Pages from-to

83-92

UT code for WoS article

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EID of the result in the Scopus database

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