[Preparation of a monoclonal antibody against polyhedrin of Ectropis obliqua nucleopolyhedrovirus].
To develop a method based on immunoreactions for detection of Ectropis obliqua Nucleopolyhedrovirus (EoNPV), the polyhedra of the virus were purified and used to immunize the mouse BALB/c. The spleen cells from the immunized mice were then fused with the myeloma cell line Sp2/0. A hybridoma cell line which can stably secrete the monoclonal antibody against EoNPV was achieved by using indirect ELISA screening and cloning methods, and was named as 7D3. Meanwhile, the polyhedrin gene was cloned from EoNPV and expressed in E. coli. Western blotting analysis showed that the monoclonal antibody prepared from 7D3 could specifically react with the recombinant polyhedrin. An indirect ELISA method based on this monoclonal antibody for detecting EoNPV in infected tea looper was developed.
Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, Zhejiang, China.
Junli
J
Du
Chuanxi
C
Zhang
Jianyu
J
Fu
Zhengxian
Z
Chen
Qiang
Q
Xiao
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D000918:Antibody Specificity; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D006825:Hybridomas / Q000378:metabolism*; D007915:Lepidoptera / Q000254:growth & development / Q000821:virology*; D051379:Mice; D008807:Mice, Inbred BALB C; D000077183:Occlusion Body Matrix Proteins; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015678:Viral Structural Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000077183:Occlusion Body Matrix Proteins; D011994:Recombinant Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus