Expression and generation of the polyclonal antibody of human group H rotavirus VP7 protein
Objective To express and block the VP7 protein of human group H rotavirus(group H rotavirus,RVH)J19,and to prepare the polyclonal adsorbent polyantibody function of VP7 for identification.Methods The VP7 protein of human H rotavirus J19 strain was expressed by baculovirus system,and the protein was blocked by affinity chromatography and hydrophobic chromatography.The VP7 polyantiserum was prepared by immunizing the VP7 protein of human H group H rotavirus J19,and the VP7 polyantiserum was identified by ELISA,Western blot(WB)and immunofluorescence.Results Soluble J19 VP7 protein was obtained.J19 VP7 rabbit polyclonal antibody was prepared.ELISA showed that VP7 polyclonal antibody had a good binding effect,binding to human RVH VP7 protein at 2 048 000-fold.WB showed that the polyclonal antibody could bind to J19 VP7 protein;Immunofluorescence assay showed that VP7 polyclonal antibody could specifically recognize human group H rotavirus J19.Conclusions The J19 VP7 protein of human H rotavirus was successfully obtained,and the prepared VP7 polyclonal antibody could recognize human H rotavirus,which provided a basis for the detection of human H rotavirus and the study of the structure and function of the viral capsid.
Human group H rotavirus(RVH)Baculovirus expression systemVP7 proteinPolyclonal antibody
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