• Applied Biological Chemistry
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• v.49 no.3
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• pp.196-201
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• 2006

Vitellogenin, which is found in the serum of female and male fishes exposed to environmental endocrine disrupter or estrogen hormone, is used as a biomarker for environmental contamination with an endocrine disrupter. In order to produce antibody against vitellogenin, a synthetic peptide for partial vitellogenin was injected into rabbits. In addition, by using ion exchange chromatography on DE-52, vitellogenin was purified from the serum of carp induced with $17{\beta}$-estradiol. Polyclonal antibody against purified vitellogenin reacted well with vitellogenin in the serum of carp induced with $17{\beta}$-estradiol and the serum of female carp, whereas polyclonal antibody against the vitellogenin peptide did not react with proteins in those samples. This may indicate that vitellogenin proteins, covalently modified largely, could not be detected by Western blotting with the polyclonal antibody against the synthetic vitellogenin peptide.

• Environmental Analysis Health and Toxicology
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• v.16 no.1
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• pp.21-28
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• 2001

A common used endpoint in bioassays testing the estrogenicity of chemicals including endocrine disruptors is the induction of egg yolk Precursor vitellogenin in male fish. Two marine fishes (Sebastes schlegeli and Paralichthys olivaceus) were exposed to the 17$\beta$-estradiol(E2) to determine the vitellogenin production. Vitellogenin was measured in fish blood using SDS-PAGE and Densimetry. Results showed that exposure to E2 caused vitellogenin in male fish. Especially, vitellogenin levels in young fish were about 4 times higher than in adult fish, which means young fish are more sensitive to E2 exposure. And plasma vitellogenin in fish increased related to E2 concentration and exposure duration.

• 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.

• The Korean Journal of Zoology
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• v.26 no.2
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• pp.83-93
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• 1983

Vitellogenin was purified from the haemolymph of female pupae during egg formation of Bombyx mori using DEAE-cellulose column and its quantitative change in various organs with age ws examined by electrophoresis and immunodiffusion. Vitellogenin is distributed in the haemolymph, fat body, and over the egg maturation and especially maintainsa constant level in the haemolymph until just before emergence, indicating that vitellogenin in released into the haemolymph at the same amount as it is taken up by oocytes during pupal period. Immunologically vitellogenin was confirmed to be in the fat body until 7 days after pupation and to undergo a drastic decline thereafter. Also, the interaction of anti-ovary proteins with haemolymph proteins showed at least 3 homogeneous proteins, indicating that other proteins as well as vitellogenin are involved in egg formation.

• Applied Biological Chemistry
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• v.49 no.3
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• pp.254-258
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• 2006

Vitellogenin has been known as a potent biomarker protein for the estrogenic activity in fish exposed to endocrine disruptors. In this study, a piezoelectric immunosensor making use of an anticarp vitellogenin antibody and an AT-cut quartz crystal microbalance as the biological component and transducer was prepared, followed by its application to the analysis of carp vitellogenin as follows. Antibody immobilization was conducted by chemisorption of a thiolated antibody with a heterobifunctional thiolation cross-linker, sulfosuccinimidyl 6-[3-(2-pyridyldithio)propionamido]hexanoate. The reaction buffer for the immunosensor system was optimized as 0.1 M sodium phosphate (pH 7.4). Concentration-dependent sensor responses were obtained in the vitellogenin concentrations ranging from 0.4864 to 486.4000 nM, with a linear correlation between vitellogenin concentration and frequency shift in double-logarithmic scale. The limit of detection of the immunosensor for carp vitellogenin was presumed as 0.4864 nM.

• Proceedings of the Korean Environmental Sciences Society Conference
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• 2001.05a
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• pp.238-239
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• 2001

The egg-yolk precursor vitellogenin(VTG) is secreted by the liver of female and male fish, in response to estrogenic compound. Carp(Cryprinus carpio) vitellogenin of one major protein is 160kDa, two minor proteins is 110kDa and 170kDa. We were induced vitellogenin by inject of 17-estradiol and purified in a two step procedure by separating it from plasma protein precipitated by 35% saturated ${(NH_4)}_2SO_4$ and then from the remainder by Mono-Q chromatography. We found major carp(Cyprinus carpio) vitellogenin band at 160kDa. Effect of different concentration of oestrogen on vitellogenin synthesis in carp(Cyprinus carpio) exposed for 4 weeks. Show differential effects on vitellogenin synthesis 7 days after treatment. Plasma vitellogenin was measured 3 times for each by an enzyme linked immunosorbent assay(ELISA). ELISA was developed for the detection of the egg yolk precursor vitellogenin in plasma of carp(Cyprinus carpio). The ELISAS performance was optimized and characterized.

• The Korean Journal of Zoology
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• v.33 no.4
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• pp.428-434
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• 1990

Uptake and accumulation of vitellogenin into ovary were traced by radiolabeled vitellogenin produced in the fat body. Vitellogenin reached the oocytes through the follicle cells and the intercellular space and penetrated the oocyte membrane via the endocytotic vesicles and then accumulated to the growing yolk granules.

• Environmental Analysis Health and Toxicology
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• v.16 no.1
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• pp.29-34
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• 2001

The estrogenic potency of di -2-ethylhexyl phthalate (DEHP) using reverse transcriptase-PCR respouse of liver vitellogenin mRNA in male African clawed frog (Xenopus laevis) was studied. Male frogs were injected with DEHP at dose of 300$\mu\textrm{g}$/kg and 300 mg/kg body weight through the dorsal lymph sac. After 4 days, using suitable pair of RT-PCR primers, vitellogenin mRNA induction in the liver was measured and DEHP showed vitellogenin mRNA induction in only the group treated with 300 mg/kg. Any significant histological abnormalities by the exposure of DEHP was not shown in both testis and liver.

• Journal of Aquaculture
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• v.9 no.1
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• pp.83-92
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• 1996

This study was conducted to compare the immunochemical properties of female-specific serum proteins (vitellogenin, VTG) and egg yolk proteins in female fusilier, Caesio diagramma. VTG of fusilier was identified and characterized by using immunochemical analysis. Two types of VTG (VTG1 and VTG2) reacted clearly with antiserum against egg proteins, were confirmed in the serum of mature female. The results of sephacryl S-300 showed that the molecular weights of VTG1 and VTG2 were 560,000 and 410,000, respectively. Yolk proteins, E2 and E3, were isolated from egg extracts, and molecular weights of them were estimated 410,000 and 170,000, respectively. The treatment of $17\beta$-estradiol ($E_2$) to males has induced the synthesis of VTG of which immunological characteristics seems to be similar to the yolk proteins. The results suggest that VTG can be synthesized in the liver by the action of $E_2$ stimulation, and incorporated into the oocytes through the blood circulation. The level of serum $E_2$ was moderately high throughout the spawning period of June. The level of serum VTG was also sustained at high in May and June. The concentration changes of serum $E_2$ and VTG were corelated to the ovarian development in female fusilier. The results indicated that $E_2$ may have some important roles for the vitellogenesis in female fusilier. Also) the VTG can be a precursor protein of yolk not only because it could be synthesized in the liver then incorporated into the oocytes but also because an egg yolk protein had the similiar molecular weights and antigenecity with VTG.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2001.05a
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• pp.132-132
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• 2001

Vitellogenin은 난황단백질의 전구체로서 생체내 발현은 에스트로젠과 에스트로젠 수용체가 반응함으로써 활성화되는 것으로 알려져 있다. 어류의 경우 간에서의 vitellogenin의 합성 및 분비는 성숙된 암컷에서만 일어날 수 있는 현상이나, 과거 여러 실험 결과를 바탕으로 수컷이나 미성숙한 암컷에서도 에스트로젠성 물질에 의해 합성이 유도되는 것으로 밝혀졌다. 따라서 수컷 또는 치어 plasma에서 vitellogenin의 측정은 외인성에스트로젠에 대한 노출을 평가할 수 있는 좋은 수단이 될 수 있다. (중략)