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Preparation of polyclonal antibody of coxsackievirus B3

Zhanfeng Jin

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Abstract

Objective To prepare polyclonal antibody against coxsackievirus B3(CVB3).Methods Specific primers which based on VP1 gene of CVB3 were designed,VP1 gene amplified by RT-PCR was cloned into pGEX-6p-1.High titer polyclonal antiserum was obtained by immunizing New Zealand rabbits with GST-VP1 fusion protein which was induced by IPTG.Then titer of the polyclonal antibody was determined by ELISA and specificity of it was tested by Western blot and indirect immunofluorescence assay.Results The titer of antiserum against CVB3 was above 1∶ 64 000,Western blot and indirect immunofluorescence analysis showed that the polyclonal antibody could bind to the CVB3 protein specifically.Conclusion High specific polyclonal antibody against CVB3 was prepared,which might be used for detection of CVB3 protein in commercial products.

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What this paper is about

Objective To prepare polyclonal antibody against coxsackievirus B3(CVB3).Methods Specific primers which based on VP1 gene of CVB3 were designed,VP1 gene amplified by RT-PCR was cloned into pGEX-6p-1.High titer polyclonal antiserum was obtained by immunizing New Zealand rabbits with GST-VP1 fusion protein which was induced by IPTG.Then titer of the polyclonal antibody was determined by ELISA and specificity of it was tested by Western blot and indirect immunofluorescence assay.Results The titer of antiserum against CVB3 was above 1∶ 64 000,Western blot and indirect immunofluorescence analysis showed that the polyclonal antibody could bind to the CVB3 protein specifically.Conclusion High specific polyclonal antibody against CVB3 was prepared,which might be used for detection of CVB3 protein in commercial products.

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Available abstract

Objective To prepare polyclonal antibody against coxsackievirus B3(CVB3).Methods Specific primers which based on VP1 gene of CVB3 were designed,VP1 gene amplified by RT-PCR was cloned into pGEX-6p-1.High titer polyclonal antiserum was obtained by immunizing New Zealand rabbits with GST-VP1 fusion protein which was induced by IPTG.Then titer of the polyclonal antibody was determined by ELISA and specificity of it was tested by Western blot and indirect immunofluorescence assay.Results The titer of antiserum against CVB3 was above 1∶ 64 000,Western blot and indirect immunofluorescence analysis showed that the polyclonal antibody could bind to the CVB3 protein specifically.Conclusion High specific polyclonal antibody against CVB3 was prepared,which might be used for detection of CVB3 protein in commercial products.

Key concepts: Polyclonal antibodies, Antiserum, Titer, Western blot, Antibody, Molecular biology, Immunofluorescence, Virology

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