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Production of a polyclonal antibody to the VP26 nucleocapsid protein of white spot syndrome virus （wssv） and its use as a biosensor

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Frontiers of Chemical Science and Engineering 2012年第2期6卷 216-223页

作者： Suchera LOYPRASERT-THANANIMIT Akrapon SALEEDANG Proespichaya KANATHARANA panote thavarungkul Wilaiwan CHOTIGEAT Center for Genomics and Bioinformatics Research Faculty of SciencePrince of Songkla UniversitySongkhla 90112Thailand Department of Molecular Biotechnology and Bioinformatics Faculty of SciencePrince of Songkla UniversitySongkhla 90112Thailand Trace Analysis and Biosensor Research Center Prince of Songkla UniversitySongkhla 90112Thailand Department of Chemistry Faculty of SciencePrince of Songkla UniversitySongkhla 90112Thailand Department of Physics Faculty of SciencePrince of Songkla UniversitySongkhla 90112Thailand

White spot syndrome virus （WSSV） is a major cause of high mortality in cultured shrimp all over the world. VP26 is one of the structural proteins of WSSV that is assumed to assist in recognizing its host and assists the viral nucleocapsid to move toward the nucleus of the host cell. The objective of this work was to produce a polyclonal antibody against VP26 and use it as a biosensor. The recombinant VP26 protein （rVP26） was produced in E. coli （BL21）, purified and used for immunizing rabbits to obtain a polyclonal antibody. Western blot analysis confirmed that the antiserum had a specific immunoreac- tivity to the VP26 of WSSV. This VP26 antiserum was immobilized onto a gold electrode for use as the sensing surface to detect WSSV under a flow injection system. The impedance change in the presence of VP26 was monitored in real time. The sensitivity linear range of 160 160000 of the biosensor was in the copies of WSSV, indicating that it is good and sensitive for analysis of WSSV. The specificity of the biosensor was supported by the observation that no impedance change was detected even at high concentrations when using Yellow Head Virus （YHV）. This biosensor may be applied to monitor the amount of WSSV in water during shrimp cultivation.

关键词： recombinant protein polyclonal antibody label-free biosensor impedance white spot syndrome virus (WSSV)

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