[Expression and assay in vitro of phage single chain antibody against nucleoprotein of influenza virus A type].
OBJECTIVE
To expression the phage single chain antibody against nucleoprotein of influenza virus A type in E. coli strain HB2151 by screening the positive clone from the phage antibody library against nucleoprotein, which will prepare for the construction of fast leak kit of Influenza virus A type.


METHODS
The positive clone ratio in the phage antibody library was enriched by three-round continual solid panning, ELISA, SDS-PAGE and Western-blot methods were used to analyze the protein secreted into the supemants and periplasmic. Affinity chromatography was used to purified the protein expressed in periplasmic and the titer was also analyzed using ELISA method.


RESULTS
10 clones were screened from96 clones and among the 10, there were 3 stronger positive with OD450nm value of 0.469, 0.582 and 0.507, respectively. The phage single chain antibody against Influenza virus A type was primary expressed in periplasmic. The ELISA results were positive even if the single chain antibody purified by affinity chromatography was diluted 4096 folds.


CONCLUSIONS
Using phage antibody library technique, phage single antibody against nucleoprotein of influenza virus A type was preliminary expressed in E. coli.
Fang
Shi-Song
SS
South China University of Technology, Guangzhou 510642, China.
Zhao
Yu-Mei
YM
He
Jian-Fan
JF
Liu
Tao
T
Liu
Jianjun
J
Zhao
Shujin
S
D000914:Antibodies, Viral / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D019590:Nucleocapsid Proteins; D019151:Peptide Library; D016133:Polymerase Chain Reaction; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000276:immunology; D057127:Single-Chain Antibodies / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D014758:Viral Core Proteins / Q000235:genetics* / Q000276:immunology
D000914:Antibodies, Viral; C048842:NP protein, Influenza A virus; D019590:Nucleocapsid Proteins; D019151:Peptide Library; D016601:RNA-Binding Proteins; D057127:Single-Chain Antibodies; D014758:Viral Core Proteins