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

Abstract

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 immunoreactivity 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 of the biosensor was in the linear range of 160–160000 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.

Key words： recombinant protein; polyclonal antibody; label-free biosensor; impedance; white spot syndrome virus (WSSV)

Acknowledgments

This work was supported by the Center for Genomics and Bioinformatics Research; Faculty of Science Research Fund, Prince of Songkhla University (PSU); the Higher Education Research Promotion and National Research University Project of Thailand, Office of the Higher Education Commission and the trace analysis and Biosensor Reasarch Center (TAB-RC). The authors also thank Dr. Brian Hodgson, Prince of Songkla University, Thailand for the English assistance with the manuscript.

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