지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
커뮤니티
연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2013.12
- 수록면
- 83 - 90 (8page)
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초록· 키워드
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Objectives Fish vitellogenin (VTG) is produced in the female liver during oogenesis through the estradiol cycle and produced in the male liver by endocrine disrupting chemicals (EDCs) such as alkylphenols. In this study, we propose that the VTG concentration in the pale chub could be detected using monoclonal antibodies and polyclonal antibodies against vitellin (Vn) in a VTG enzyme-linked immunosorbent assay (ELISA) system.
Methods Monoclonal antibodies and polyclonal antibodies were produced using the Vn extracted from the matured ovum of the ovary. The VTG was extracted from the plasma of the male pale chub. The Vn and VTG were confirmed by measuring the molecular weight of their proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDSPAGE) and the specificity of the antibodies was checked through western blotting methods. The assay system was validated with respect to optimal assay concentrations, specificity, recovery, and intra- and inter-assay variations.
Results The Vn consisted of two protein bands with apparent molecular weights of 64 and 37 kDa. The SDS-PAGE indicated protein weights of 146 and 77 kDa in the VTG. The assay range was 15.6 ng/mL to 2,000 ng/mL, and the value of the intra- and inter-assay variations were within 10.0% and 14.7%, respectively. The recovery rate was 99.5±5.5%.
Conclusions A sandwich ELISA was developed that could be used to qualify the VTG of pale chub in screening for EDCs. Pale chub is an ideal species for observing estrogen activity in the environment because of its extensive habitat and extensive food chain. The ELISA developed here would be more favorable than those for other species for determining the effect of long-term food chain accumulation of EDCs in aquatic environments.
Methods Monoclonal antibodies and polyclonal antibodies were produced using the Vn extracted from the matured ovum of the ovary. The VTG was extracted from the plasma of the male pale chub. The Vn and VTG were confirmed by measuring the molecular weight of their proteins using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDSPAGE) and the specificity of the antibodies was checked through western blotting methods. The assay system was validated with respect to optimal assay concentrations, specificity, recovery, and intra- and inter-assay variations.
Results The Vn consisted of two protein bands with apparent molecular weights of 64 and 37 kDa. The SDS-PAGE indicated protein weights of 146 and 77 kDa in the VTG. The assay range was 15.6 ng/mL to 2,000 ng/mL, and the value of the intra- and inter-assay variations were within 10.0% and 14.7%, respectively. The recovery rate was 99.5±5.5%.
Conclusions A sandwich ELISA was developed that could be used to qualify the VTG of pale chub in screening for EDCs. Pale chub is an ideal species for observing estrogen activity in the environment because of its extensive habitat and extensive food chain. The ELISA developed here would be more favorable than those for other species for determining the effect of long-term food chain accumulation of EDCs in aquatic environments.
목차
Introduction
Materials and Methods
Results
Discussion
References
참고문헌 (24)
참고문헌 신청
Ackermann GE / 2002 / Effects of longterm nonylphenol exposure on gonadal development and biomarkers of estrogenicity in juvenile rainbow trout Oncorhynchus mykiss / Aquat Toxicol 60(3-4):203~221
Heemken OP / 2001 / The occurrence of xenoestrogens in the Elbe river and the North Sea / Chemosphere 45(3):245~259
Hutchinson TH / 2002 / Ecological risk assessment and testing for endocrine disruption in the aquatic environment / Toxicology 181(182):383~387
Denslow ND / 1999 / Vitellogenin as a biomarker of exposure for estrogen or estrogen mimics / Ecotoxicology 8(5):385~398
Bowman CJ / 2000 / Estrogen- induced vitellogenin mRNA and protein in sheepshead minnow (Cyprinodon variegatus) / Gen Comp Endocrinol 120(3):300~313
Heemken OP / 2001 / The occurrence of xenoestrogens in the Elbe river and the North Sea / Chemosphere 45(3):245~259
Hutchinson TH / 2002 / Ecological risk assessment and testing for endocrine disruption in the aquatic environment / Toxicology 181(182):383~387
Denslow ND / 1999 / Vitellogenin as a biomarker of exposure for estrogen or estrogen mimics / Ecotoxicology 8(5):385~398
Bowman CJ / 2000 / Estrogen- induced vitellogenin mRNA and protein in sheepshead minnow (Cyprinodon variegatus) / Gen Comp Endocrinol 120(3):300~313
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UCI(KEPA) : I410-ECN-0101-2015-530-001447498