[Preparation and partial characterization of monoclonal antibodies specific for nuclear protein of avian influenza virus].
OBJECTIVE
To prepare the monoclonal antibodies (mAbs) specific for nuclear protein (NP) of avian influenza virus (AIV) and identify their biological properties.


METHODS
BALB/c mice were immunized with AIV (formaldehyde-inactivated AIV H9N2, Triton X-100-lysed H9N2 and AIV NP expressed in E.coli, respectively). Hybridoma cell lines secreting anti-AIV NP mAbs were developed through cell fusion, screening and cloning. The mAb's titer was determined by indirect ELISA. Specificity of mAbs was identified by cross-reaction test and indirect immuno-fluorescence assay (IFA).


RESULTS
6 hybridoma cell lines secreting anti-AIV NP mAbs were obtained, designated 4F4, 1C3, 1G11, 1C2, 1D10 and 2F7. ELISA detection showed that the titers of two mAbs (1G11 and 1D10) out of 6 mAbs were the highest (2(-13) and 2(-14), respectively) and their specificity was also better than that of the others, confirmed by cross-reaction test and IFA.


CONCLUSIONS
In this study 6 mAbs against AIV NP were obtained. The mAbs 1G11 and 1D10 perform the best in titer and specificity. This work paves the way for AIV study and development of method for rapid detection of AIV.
Liu
Fang-Fang
FF
Animal Pathogenic Microbiology of National Key Laboratory of Agricultural Microbiology, Centre China Agriculural University, Wuhan 430070, China.
Jin
Mei-lin
ML
Zhang
An-ding
AD
Dan
Han-bing
HB
Chen
Huan-chun
HC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis / Q000276:immunology*; D000918:Antibody Specificity; D001717:Birds / Q000821:virology; D002460:Cell Line; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas / Q000378:metabolism; D053127:Influenza A Virus, H9N2 Subtype / Q000276:immunology* / Q000302:isolation & purification; D005585:Influenza in Birds / Q000821:virology; D009687:Nuclear Proteins / Q000276:immunology*; D014764:Viral Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D009687:Nuclear Proteins; D014764:Viral Proteins