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- W2381160546 abstract "Objective To generate rabbit polyclonal antibody against Kaposi's sarcoma-associated herpesvirus( KSHV) encoding peptides of viral interleukin-6( v IL-6),identify the specificity of the prepared polyclonal v IL-6 antibody by constructing the recombinant plasmid,and detect the native viral protein using the prepared polyclonal v IL-6 antibody. Methods After the bioinformatics analysis for the conformational antigenic epitopes of the v IL-6,three candidate polypeptides of v IL-6 were designed and synthesized. New Zealand white rabbits were immunized with the synthetic peptides to generate polyclonal antibodies against v IL-6. Enzyme-linked immunosorbent assay( ELISA) was performed to characterize the titers of the prepared polyclonal antibodies. The recombinant plasmid,p HAGE-v IL-6,was constructed to transfect 293 T and EA. hy926 cells to obtain v IL-6-His fusion protein,which was identified by western blot. The native viral protein in KSHV-infected primary effusion lymphoma( PEL) cells was recognized using the polyclonal antibodies against v IL-6 by western blot and immunofluorescence assay( IFA). Results The titer of the polyclonal antibodies against v IL-6 prepared with the third antigenic polypeptide was greater than 8 000 based on ELISA and could react specifically with the v IL-6-His fusion protein in 293 T and EA. hy926 cells as well as the native viral protein in PEL cells. Conclusion We successfully prepared the polyclonal antibody against v IL-6 to react with fusion protein and native viral protein by using the synthesized peptides as hapten,which should be available for detecting recombinant and native v IL-6 protein." @default.
- W2381160546 created "2016-06-24" @default.
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- W2381160546 date "2015-01-01" @default.
- W2381160546 modified "2023-09-23" @default.
- W2381160546 title "Generation and characterization of polyclonal antibody against synthetic Kaposi' s sarcoma-associated herpesvirus-encoding peptides of viral interleukin-6 and its application" @default.
- W2381160546 hasPublicationYear "2015" @default.
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