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

被引:0
作者
Suchera Loyprasert-Thananimit
Akrapon Saleedang
Proespichaya Kanatharana
Panote Thavarungkul
Wilaiwan Chotigeat
机构
[1] Center for Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University
[2] Department of Molecular Biotechnology and Bioinformatics, Faculty of Science, Prince of Songkla University
[3] Trace Analysis and Biosensor Research Center, Prince of Songkla University
[4] Department of Chemistry, Faculty of Science, Prince of Songkla University
[5] Department of Physics, Faculty of Science, Prince of Songkla University
关键词
impedance; label-free biosensor; polyclonal antibody; recombinant protein; white spot syndrome virus (WSSV);
D O I
10.1007/s11705-012-1289-y
中图分类号
学科分类号
摘要
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. © 2012 Higher Education Press and Springer-Verlag Berlin Heidelberg.
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页码:216 / 223
页数:7
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