Monoclonal antibodies produced against VP3 of a novel mud crab dicistrovirus.
Mud crab dicistrovirus (MCDV) is a recently identified single-stranded positive RNA virus, which causes serious economic loss. The three structural proteins of MCDV were separated by SDS-PAGE. In this study, a recombinant MCDV-VP3 was successfully expressed by an Escherichia coli expression system. After immunization and cell fusion, three mouse hybridomas (1G6H9, 1E7B8, 4B6E10) producing MAbs against MCDV-VP3 were established. The MAbs obtained were fully characterized using indirect ELISA and Western blot analysis. The ELISA results showed that the titers of MAbs were in the range of 1:3.200×10(3) in culture fluids and 1:2.048×10(5) in ascitic fluids. Using Western blot analyses, we observed the specific characteristic band that defines VP3. It demonstrated that all the MAbs were directed against MCDV-VP3. The results of immunogold transmission electron microscopy (IEM) suggested that MCDV-VP3 is the capsid protein of MCDV. The preparation of MAbs specific to the structure protein of MCDV would be useful for studying the function of the structure proteins and the mechanism of infection and pathogenesis of MCDV.
State Key Laboratory for Biocontrol, School of Life Sciences, Sun Yat-Sen Zhongshan University, Guangzhou, People's Republic of China.
Rui
R
Zhang
Jianguo
J
He
Hongjun
H
Su
Chuanfu
C
Dong
Zhixun
Z
Guo
Shaoping
S
Weng
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D000918:Antibody Specificity; D015153:Blotting, Western; D003386:Brachyura / Q000821:virology*; D056629:Dicistroviridae / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D014758:Viral Core Proteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D014758:Viral Core Proteins