A monoclonal antibody against Lates calcarifer vitellogenin and a competitive ELISA to evaluate vitellogenin induction after exposure to xenoestrogen
| Autor: | Sutin Kingtong, Nanthika Khongchareonporn, Witchuda Prasatkaew, Phochit Nanthanawat |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Male
Environmental Engineering food.ingredient medicine.drug_class 0211 other engineering and technologies Enzyme-Linked Immunosorbent Assay 02 engineering and technology Endocrine Disruptors 010501 environmental sciences Monoclonal antibody 01 natural sciences Vitellogenins chemistry.chemical_compound Bass (fish) Vitellogenin food Phenols Benzo(a)pyrene medicine Animals Environmental Chemistry Grouper Sea bass 0105 earth and related environmental sciences General Environmental Science 021110 strategic defence & security studies biology Antibodies Monoclonal Estrogens General Medicine Thailand biology.organism_classification Molecular biology Nonylphenol Xenoestrogen chemistry biology.protein Bass Antibody Water Pollutants Chemical |
| Zdroj: | Journal of Environmental Sciences. 75:325-333 |
| ISSN: | 1001-0742 |
| DOI: | 10.1016/j.jes.2018.05.001 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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