Purification, Partial Characterization, and Immunoassay of Vitellogenin from Marbled Sole (Limanda yokohamae) KCI

Title
Purification, Partial Characterization, and Immunoassay of Vitellogenin from Marbled Sole (Limanda yokohamae)
Author(s)
김대중; 민광식; 김윤; 정지현; 한창희; 안철민; 지영주
KIOST Author(s)
Jung, Jee Hyun(정지현)
Publication Year
2005-12
Abstract
Vitellogenin (VTG) was purified from the blood plasma of estradiol-17 (E2)-treated male marbled sole (Limanda yokohamae) using gel filtration and anion exchange chromatography. The purity of the marbled sole VTG (msVTG) was confirmed by polyacrylamide gel electrophoresis (SDS-PAGE) and N-terminal amino acid sequencing. The purified msVTG was used to produce monoclonal and polyclonal antibodies in mice and rabbits, respectively, and the specificity of the polyclonal antisera for msVTG was confirmed by Western blot analysis. The antibodies cross-reacted with a protein of molecular mass approximately 160 kDa in the plasma samples of mature female marbled sole. No cross-reactivity was observed with the plasma of male fish. A direct non-competitive sandwich enzyme-linked immunosorbent assay (ELISA) was developed using the monoclonal anti-msVTG and polyclonal anti-msVTG antibodies, with purified msVTG as the standard protein. The values of the intra- and inter-assay variations were within the ranges of 8.1-9.8% and 8.5-12.2%, respectively. The sensitivity was about 0.3 ng/mL. Serial dilutions of plasma from mature female sole reacted with the msVTG-antibodies in the sandwich ELISA, whereas the plasma from male fish did not. The results indicate that the maturation status of female marbled sole can be identified using a sandwich ELISA for msVTG.
ISSN
2234-1749
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/4994
Bibliographic Citation
Fisheries and Aquatic Sciences, v.8, no.4, pp.213 - 219, 2005
Publisher
한국수산과학회
Keywords
Marbled sole; Limanda yokohamae; vitellogenin; monoclonal antibody; polyclonal antibody; sandwich ELISA
Type
Article
Publisher
한국수산과학회
Related Researcher
Research Interests

Toxicology,Fish physiology,Marine environment risk assessment,독성학,어류생리학,해양환경 위해성평가

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