Rockfish (Sebastes schlegeli) Vitellogenin: Purification, Characterization and Development of Sandwich ELISA System KCI

Title
Rockfish (Sebastes schlegeli) Vitellogenin: Purification, Characterization and Development of Sandwich ELISA System
Author(s)
정지현; 김대중; 한창희
KIOST Author(s)
Jung, Jee Hyun(정지현)
Publication Year
2004-09
Abstract
Vitellogenin (VTG) was purified from serum of estradiol-17β-treated rockfish (Sebastes schlegeli) by precipitation with EDTA-Mg2+and ammonium sulfate and two step chromatography (anion exchange chromatography and gel permeation chromatography) was performed on FPLC system. Rockfish VTG (rfVTG) was characterized and its properties were determined. The monomers have apparent, molecular mass of about 188kDa as indicated by SDS-PAGE. Amino acid composition analysis of rfVTG was similar to VTG from other oviparous teleosts. Cysteine and lysine were present at relatively high level. Leucine was present at relatively lower level than in other species. The N-terminal amino acid sequence was evaluated to identify rfVTG. Western blot analysis using an antibody against the purified VTG showed that the antibody reacted with both plasma of estradiol-17β treated male and purified VTG, whereas there was no reaction with male serum of the control. An ELISA was developed using monoclonal and polyclonal antibodies against rfVTG. The assay range was 3.2ng/mL and 1,000ng/mL and the value of the intra and inter assay variations were within 9.7% and 11.2%, respectively. Recovery rate was 96.8%. The sandwich ELISA could be useful for the detection of VTG and could be good for screening of estrogenic compounds.
ISSN
2234-1749
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/5220
Bibliographic Citation
Fisheries and Aquatic Sciences, v.7, no.3, pp.99 - 108, 2004
Publisher
한국수산과학회
Keywords
ELISA; Rockfish; Sebastes schlegeli; Vitellogenin (VTG); Monoclonal antibody; Polyclonal antibody
Type
Article
Publisher
한국수산과학회
Related Researcher
Research Interests

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

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