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Monoclonal antibody against recombinant Fasciola gigantica cathepsin L1H could detect juvenile and adult cathepsin Ls of Fasciola gigantica

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Abstract

Cathepsin Ls (CatLs), the major cysteine protease secreted by Fasciola spp., are important for parasite digestion and tissue invasion. Fasciola gigantica cathepsin L1H (FgCatL1H) is the isotype expressed in the early stages for migration and invasion. In the present study, a monoclonal antibody (MoAb) against recombinant F. gigantica cathepsin L1H (rFgCatL1H) was produced by hybridoma technique using spleen cells from BALB/c mice immunized with recombinant proFgCatL1H (rproFgCatL1H). This MoAb is an immunoglobulin (Ig)G1 with κ light chain isotype. The MoAb reacted specifically with rproFgCatL1H, the native FgCatL1H at a molecular weight (MW) 38 to 48 kDa in the extract of whole body (WB) of metacercariae and newly excysted juvenile (NEJ) and cross-reacted with rFgCatL1 and native FgCatLs at MW 25 to 28 kDa in WB of 2- and 4-week-old juveniles, adult, and adult excretory–secretory (ES) fractions by immunoblotting and indirect ELISA. It did not cross-react with antigens in WB fractions from other parasites, including Gigantocotyle explanatum, Paramphistomum cervi, Gastrothylax crumenifer, Eurytrema pancreaticum, Setaria labiato-papillosa, and Fischoederius cobboldi. By immunolocalization, MoAb against rFgCatL1H reacted with the native protein in the gut of metacercariae and NEJ and also cross-reacted with CatL1 in 2- and 4-week-old juveniles and adult F. gigantica. Therefore, FgCatL1H and its MoAb may be used for immunodiagnosis of both early and late fasciolosis in ruminants and humans.

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Acknowledgments

This research was supported by grants from the Faculty of Science, Mahidol University to Suda Riengrojpitak and Prasert Sobhon.

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Authors and Affiliations

  1. Department of Pathobiology, Faculty of Science, Mahidol University, Rama VI Rd, Ratchathewi, Bangkok, 10400, Thailand

    Sirima Wongwairot, Narin Changklungmoa, Wipaphorn Jaikua & Suda Riengrojpitak

  2. Faculty of Allied Health Sciences, Burapha University, Long-Hard Bangsaen Rd, Saen Sook Sub-district, Mueang District, Chonburi, 20131, Thailand

    Pornanan Kueakhai

  3. Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd, Ratchathewi, Bangkok, 10400, Thailand

    Krai Meemon, Sineenart Songkoomkrong & Prasert Sobhon

  4. Faculty of Physical Therapy, Mahidol University, Putthamonthon Sai 4 Rd, Salaya, Putthamonthon, Nakhon Pathom, 73170, Thailand

    Veerawat Sansri

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  1. Sirima Wongwairot
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  2. Pornanan Kueakhai
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  3. Narin Changklungmoa
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  6. Krai Meemon
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  7. Sineenart Songkoomkrong
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  8. Suda Riengrojpitak
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  9. Prasert Sobhon
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Correspondence to Suda Riengrojpitak.

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Wongwairot, S., Kueakhai, P., Changklungmoa, N. et al. Monoclonal antibody against recombinant Fasciola gigantica cathepsin L1H could detect juvenile and adult cathepsin Ls of Fasciola gigantica . Parasitol Res 114, 133–140 (2015). https://doi.org/10.1007/s00436-014-4170-8

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  • DOI: https://doi.org/10.1007/s00436-014-4170-8

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