• Journal of Microbiology and Biotechnology
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• v.17 no.9
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• pp.1445-1451
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• 2007

A sensitive optical waveguide lightmode spectroscopy-based immunosensor was developed to detect vitellogenin in seawater flatfish (Paralichthys olivaceus). For this purpose, anion-exchange column chromatography with DE-52 resin was used to purify flatfish vitellogenin from flatfish serum containing vitellogenin that had been induced using an intraperitoneal $17{\beta}$-estradiol injection. The anti-flatfish vitellogenin antibody used as the biological component of the above immunosensor was prepared using the purified flatfish vitellogenin. The change in the incoupling angle according to the complexation between the flatfish vitellogenin and its antibody, immobilized over an optical grating coupler sensor chip, was measured to calculate the sensor response. The immunosensor was quite specific to flatfish vitellogenin binding, based on no sensor response in the case of bovine serum albumin immobilization. When plotted using double-logarithmic scales, the sensor responses increased linearly in flatfish vitellogenin concentrations of 0.00675-67.5 nM, with a detection limit of 0.0675 nM. The reusability during seven repetitive measurements was reasonably fair for the preliminary screening of flatfish vitellogenin.

• Journal of Applied Biological Chemistry
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• v.52 no.4
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• pp.170-173
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• 2009

Vitellogenin (Vtg) is found in the serum of both female and male fish that have been exposed to environmental endocrine disrupters or estrogen hormones, and is used as a biomarker for such contamination. In our current study antibodies raised against the purified flatfish Vtg were tested for their reactivity on immunoblots to flatfish and carp Vtg, and also to a Vtg protein fragment produced in E. coli. Polyclonal antibodies raised against purified flatfish Vtg reacted well with Vtg in the serum of flatfish and carp induced with $17{\beta}$-estradiol, but not with the Vtg protein fragment produced in E. coli.