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Cellosaurus publication CLPUB00400

Publication number CLPUB00400
Authors Nishikawa T., Maruo N., Tatarazako N., Shiraishi H., Morita M.
Title Establishment and characterization of medaka vitellogenin monoclonal antibodies.
Citation (In) 4th annual meeting of Japan Society of Endocrine Disrupters Research; pp.PB-61; Tsukuba (2001)
Web pages https://www.env.go.jp/chemi/end/sympo/2001/report/pdf_abs/pb/PB_61.pdf
Abstract We established Medaka (Oryzias latipes) vitellogenin (VTG) monoclonal antibodies for ELISA application. Medaka Was fed by 1 mg 17 beta- estradiol/g tetrafin for seven days to collect ascite fluids containing VTG. After purification of VTG using POROS-HQ column chromatography, VTG in PBS was mixed with equal volume of Freund's adjuvant and Mice (6 weeks BALB/c females) were immunized 3 times per every 14 days. Spleen cells from immunized mouse and myeloma cells were fused using polyethylene glycol, then hybridoma cells were subjected to HAT selection. Hybridoma producing monoclonal antibodies specific to Medaka VTG were selected and four clones (MVP25, MVP47, MVP49, MVP51) were established, of which these four monoclonal antibodies did not show any reactivity to male blood and whole body and liver homogenates. Western blot analysis revealed that each of these antibodies recognized lipovitellin heavy chain (120 kDa protein existed in VTG N-terminal region) as an epitope. It was possible to construct sandwich ELISA system for Medaka VTG using MVP25-MVP47 or MVP47- MVP49, but not MVP25-MVP49, indicating MVP25 and MVP49 bind the same or near epitope.
Cell lines CVCL_U131; MVP25
CVCL_U132; MVP47
CVCL_U133; MVP49
CVCL_U134; MVP51