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A monoclonal antibody against Lates calcarifer vitellogenin and a competitive ELISA to evaluate vitellogenin induction after exposure to xenoestrogen

Prasatkaew, Witchuda, Nanthanawat, Phochit, Khongchareonporn, Nanthika, Kingtong, Sutin
Journal of environmental sciences (China) 2019 v.75 pp. 325-333
Epinephelus, Lates calcarifer, Plectropomus, antibody formation, aquatic environment, bass, benzo(a)pyrene, blood, cadmium chloride, cross reaction, endocrine-disrupting chemicals, enzyme-linked immunosorbent assay, estradiol, hybridomas, hydroxyapatite, intramuscular injection, liquid chromatography, mice, monitoring, monoclonal antibodies, nonylphenols, polyacrylamide gel electrophoresis, staining, tandem mass spectrometry, vitellogenin, Thailand
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.