| CELER, Vladimír, Dalibor BLAZEK, Iva NAVRÁTILOVÁ and Petr SKLÁDAL. Generation and characterization of single-chain antibody fragments specific against transmembrane envelope glycoprotein gp46 of Maedi-visna Virus. Journal of Virological Methods. Amsterdam: Elsevier, 2004, vol. 115, No 1, p. 83-92. ISSN 0166-0934. |
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@article{555471,
author = {Celer, Vladimír and Blazek, Dalibor and Navrátilová, Iva and Skládal, Petr},
article_location = {Amsterdam},
article_number = {1},
keywords = {Recombinant antibody; Spreeta; SPR; Kinetics},
language = {eng},
issn = {0166-0934},
journal = {Journal of Virological Methods},
title = {Generation and characterization of single-chain antibody fragments specific against transmembrane envelope glycoprotein gp46 of Maedi-visna Virus},
volume = {115},
year = {2004}
}
TY - JOUR
ID - 555471
AU - Celer, Vladimír - Blazek, Dalibor - Navrátilová, Iva - Skládal, Petr
PY - 2004
TI - Generation and characterization of single-chain antibody fragments specific against transmembrane envelope glycoprotein gp46 of Maedi-visna Virus
JF - Journal of Virological Methods
VL - 115
IS - 1
SP - 83-92
EP - 83-92
PB - Elsevier
SN - 01660934
KW - Recombinant antibody
KW - Spreeta
KW - SPR
KW - Kinetics
N2 - 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.
ER -
CELER, Vladimír, Dalibor BLAZEK, Iva NAVRÁTILOVÁ and Petr SKLÁDAL. Generation and characterization of single-chain antibody fragments specific against transmembrane envelope glycoprotein gp46 of Maedi-visna Virus. \textit{Journal of Virological Methods}. Amsterdam: Elsevier, 2004, vol.~115, No~1, p.~83-92. ISSN~0166-0934.
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