Detailed Information on Publication Record
2004
Generation and characterization of single-chain antibody fragments specific against transmembrane envelope glycoprotein gp46 of Maedi-visna Virus
CELER, Vladimír, Dalibor BLAZEK, Iva NAVRÁTILOVÁ and Petr SKLÁDALBasic information
Original name
Generation and characterization of single-chain antibody fragments specific against transmembrane envelope glycoprotein gp46 of Maedi-visna Virus
Name in Czech
Produkce a charakterisace scFv protilátek specifických proti transmembránovému obalovému glykoproteinu viru Maedi-visna
Authors
CELER, Vladimír (203 Czech Republic), Dalibor BLAZEK (203 Czech Republic), Iva NAVRÁTILOVÁ (203 Czech Republic) and Petr SKLÁDAL (203 Czech Republic, guarantor)
Edition
Journal of Virological Methods, Amsterdam, Elsevier, 2004, 0166-0934
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10600 1.6 Biological sciences
Country of publisher
Czech Republic
Confidentiality degree
není předmětem státního či obchodního tajemství
Impact factor
Impact factor: 1.729
RIV identification code
RIV/00216224:14310/04:00010104
Organization unit
Faculty of Science
UT WoS
000187795500010
Keywords in English
Recombinant antibody; Spreeta; SPR; Kinetics
Tags
Změněno: 18/1/2005 10:19, prof. RNDr. Petr Skládal, CSc.
V originále
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 scFvs for expression as intrabodies in intracellular immunization against virus maedi-visna infection and for the diagnosis of this virus is discussed.
In Czech
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 proteinu gp46. Protilátky ve formě scFv byly přečištěny metaloafinitní chromatografií a charakterisovány pomocí biosensoru na bázi povrchové resonance plasmonů Spreeta.
Links
| OC 518.30, research and development project |
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