本文主要研究内容
作者（2019）在《A monoclonal antibody against Lates calcarifer vitellogenin and a competitive ELISA to evaluate vitellogenin induction after exposure to xenoestrogen》一文中研究指出：A monoclonal antibody specific to sea bass（Lates calcarifer） vitellogenin（VTG） was developed,for use as a tool for monitoring endocrine disrupting chemicals（EDCs）. VTG was induced in sea bass by intramuscular injection of 17β-estradiol（E2: 2 mg/kg） every three days. Blood was collected three days after the last injection. Plasma VTG was then purified by chromatography in hydroxyapatite and a sephacryl-S300 column. Characterizations of purified VTG were done by phospholipoglycoprotein staining on a native-PAGE with confirmation by mass spectrometry（LC-MS/MS）. Antibody was raised in mice by injection of purified VTG. After monoclonal antibody production, the hybridoma clone No. 41（MAb-sea bass VTG 41）was selected and developed for quantification of VTG by competitive enzyme-linked immunosorbent assay（ELISA）. The ELISA method was sensitive with a detection limit of VTG 40 ng/mL. MAb-sea bass VTG 41 was specific to VTG from E2-treated sea bass and others EDCs（Nonylphenol, Benzo[a]pyrene and CdCl2）. Moreover, cross-reactivity was also found in E2-treated coral grouper（Epinephelus corallicola）. The ELISA method obtained from this work can be further applied for the assessment of EDCs in Thailand and Southeast Asia’s aquatic environment.
Abstract
A monoclonal antibody specific to sea bass（Lates calcarifer） vitellogenin（VTG） was developed,for use as a tool for monitoring endocrine disrupting chemicals（EDCs）. VTG was induced in sea bass by intramuscular injection of 17β-estradiol（E2: 2 mg/kg） every three days. Blood was collected three days after the last injection. Plasma VTG was then purified by chromatography in hydroxyapatite and a sephacryl-S300 column. Characterizations of purified VTG were done by phospholipoglycoprotein staining on a native-PAGE with confirmation by mass spectrometry（LC-MS/MS）. Antibody was raised in mice by injection of purified VTG. After monoclonal antibody production, the hybridoma clone No. 41（MAb-sea bass VTG 41）was selected and developed for quantification of VTG by competitive enzyme-linked immunosorbent assay（ELISA）. The ELISA method was sensitive with a detection limit of VTG 40 ng/mL. MAb-sea bass VTG 41 was specific to VTG from E2-treated sea bass and others EDCs（Nonylphenol, Benzo[a]pyrene and CdCl2）. Moreover, cross-reactivity was also found in E2-treated coral grouper（Epinephelus corallicola）. The ELISA method obtained from this work can be further applied for the assessment of EDCs in Thailand and Southeast Asia’s aquatic environment.
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