title: Production and characterization of a monoclonal antibody against recombinant cathepsin L1 of Fasciola gigantica.
abstract: Monoclonal antibodies (MoAbs) against a recombinant cathepsin L1 of Fasciola gigantica (rFgCatL1) were produced in vitro by fusion of BALB/c mice spleen cells immunized with rFgCatL1 and mouse myeloma cells. Reactivity and specificity of these MoAbs were evaluated by indirect ELISA and immunoblotting techniques. Seven MoAb clones were selected from the stable hybridoma clones, namely 1E10, 1F5, 3D11, 4B10, 4D3, 4E3 and 5E7. Clones 1E10, 1F5 and 3D11 were IgM, whereas clones 4B10, 4D3, 4E3 and 5E7 were IgG1. All MoAbs had kappa light chain isotypes. All MoAbs reacted with rCatL1 at molecular weight (MW) 30kDa and with the native CatL1 at MW 27kDa in whole body (WB) extracts of metacercariae (Met), newly excysted juveniles (NEJ), 1, 3, 5-week-old juveniles (Ju), adult WB and adult excretory-secretory (ES) fractions, but not with adult tegumental antigens (TA). All of these MoAbs showed no cross-reactions with antigens of other parasites commonly found in ruminants and human, including Paramphistomum cervi, Eurytrema pancreaticum, Gigantocotyle explanatum, Schistosoma spindale, Schistosoma mansoni, Moniezia benedeni, Avitellina centripunctata, Trichuris sp., Haemonchus placei and Setaria labiato-papillosa. Localization of CatL1 in each developmental stages of F. gigantica by immunoperoxidase technique, using these MoAbs as probes, indicated that CatL1 was present at high concentration in the caecal epithelium and caecal lumen of metacercariae, NEJ, 1, 3, 5-week-old juveniles and adult fluke. This finding indicated that CatL1 is a copiously expressed parasite protein that is released into the ES, thus CatL1 and its MoAb could be a good candidate for immunodiagnosis of fasciolosis in ruminant and human.
authors.affiliation: Division of Agricultural Science, Mahidol University, Kanchanaburi Campus, Saiyok, Kanchanaburi 71150, Thailand; Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd., Bangkok 10400, Thailand. Electronic address: panat.anu@mahidol.ac.th.
authors.forename: Panat
authors.initials: P
authors.lastname: Anuracpreeda
authors.affiliation: Division of Agricultural Science, Mahidol University, Kanchanaburi Campus, Saiyok, Kanchanaburi 71150, Thailand.
authors.forename: Thippawan
authors.initials: T
authors.lastname: Srirakam
authors.affiliation: Division of Agricultural Science, Mahidol University, Kanchanaburi Campus, Saiyok, Kanchanaburi 71150, Thailand.
authors.forename: Sudarat
authors.initials: S
authors.lastname: Pandonlan
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd., Bangkok 10400, Thailand.
authors.forename: Narin
authors.initials: N
authors.lastname: Changklungmoa
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd., Bangkok 10400, Thailand; Mahidol University, Nakhonsawan Campus, Nakhonsawan 60130, Thailand.
authors.forename: Charoonroj
authors.initials: C
authors.lastname: Chotwiwatthanakun
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd., Bangkok 10400, Thailand; Mahidol University, Nakhonsawan Campus, Nakhonsawan 60130, Thailand.
authors.forename: Yotsawan
authors.initials: Y
authors.lastname: Tinikul
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd., Bangkok 10400, Thailand; Mahidol University, Nakhonsawan Campus, Nakhonsawan 60130, Thailand.
authors.forename: Jaruwan
authors.initials: J
authors.lastname: Poljaroen
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd., Bangkok 10400, Thailand.
authors.forename: Krai
authors.initials: K
authors.lastname: Meemon
authors.affiliation: Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd., Bangkok 10400, Thailand.
authors.forename: Prasert
authors.initials: P
authors.lastname: Sobhon
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D002403:Cathepsins / Q000276:immunology* / Q000378:metabolism; D005209:Fasciola / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D051379:Mice; D011994:Recombinant Proteins / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D002403:Cathepsins
keywords: Cathepsin L1; Cross-reaction; Fasciola gigantica; Immunolocalization; Monoclonal antibody