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- Title
Preparation and identification of duck hepatitis B virus core protein monoclonal antibody.
- Authors
WANG Ya-wen; LIU Zheng-wen; ZHANG Lin; FENG Ai; WANG Wei; WANG Quan-ying; YANG Guang-xiao; HUI Ling-yun
- Abstract
Objective To prepare and identify monoclonal antibodies against duck hepatitis B virus (DHBV) core protein. Methods DHBV core gene was amplified by PCR to construct a recombinant plasmid pET28a(+)/ DHBV core. The recombinant plasmid was expressed in E. coli BL21 induced by IPTG and DHBV core protein purified by Ni2+ affinity chromatography was used to immunize 8-week-old female Balb/c Mus musculson. Then after fusion between spleen cells stimulated and SP2/0 myeloma cell, the hybridoma clone against DHBV core protein was obtained by limiting-dilution method and ELISA assay. A large number of anybodies were induced in vivo. The speaificity of the antibodies was identified by ELISA assay and immunohistochemistry. Results Recombinant plasmid pET28a(+)/DHBV core was expreised succeisfully in E.coli BL21. Two hybridoma clones secreting antibody against DHBV core protein, named 2D7 and 5G10, were obtained. A novel murine monoclonal aniibody (5G10) was harvetted and it was specific to DHBV core protein. Isotype analysis showed that MAb 5G10 belonged to IgG2a subclass with k light chain. Conclusion Monoclonal antibodies against DHBV core protein have been prepared and obtained in this study, which lays the foundation for research on the function of DHBV core protein, development of diagnostic reagents and anti-viral treatment.
- Subjects
HEPATITIS B virus; MONOCLONAL antibodies; VIRAL proteins; DUCKS; RECOMBINANT microorganisms; POLYMERASE chain reaction
- Publication
Journal of Xi'an Jiaotong University (Medical Sciences), 2013, Vol 34, Issue 6, p768
- ISSN
1671-8259
- Publication type
Article
- DOI
10.7652/jdyxb201306014