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For clinical application of antibodies as a positron emission tomography (<jats:styled-content style=\"fixed-case\">PET<\/jats:styled-content>) imaging reagent, a human shortened antibody is essential both for avoiding redundant immune responses and for providing rapid imaging. Therefore, we cloned a single\u2010chain fragment of variable regions (scFv) from a human\u2010derived gene sequence. This was achieved through the construction of a na\u00efve phage library derived from human tonsil lymphocytes. Using a column with human recombinant <jats:styled-content style=\"fixed-case\">MSLN<\/jats:styled-content>, we carried out bio\u2010panning of phage\u2010variants by colony formation. We first obtained 120 clones that were subjected to selection in an <jats:styled-content style=\"fixed-case\">ELISA<\/jats:styled-content> using human recombinant <jats:styled-content style=\"fixed-case\">MSLN<\/jats:styled-content> as a solid phase antigen, and 15 phage clones of scFv with a different sequence were selected and investigated by flow cytometry (<jats:styled-content style=\"fixed-case\">FCM<\/jats:styled-content>). Then, six variants were selected and the individual scFv gene was synthesized in the <jats:styled-content style=\"fixed-case\">V<jats:sub>L<\/jats:sub><\/jats:styled-content> and <jats:styled-content style=\"fixed-case\">V<jats:sub>H<\/jats:sub><\/jats:styled-content> domains and expressed in Chinese hamster ovary cells. Mammalian cell\u2010derived human\u2010origin scFv clones were analyzed by <jats:styled-content style=\"fixed-case\">FCM<\/jats:styled-content> again, and one <jats:styled-content style=\"fixed-case\">MSLN<\/jats:styled-content> highly specific scFv clone was established. <jats:styled-content style=\"fixed-case\">PET<\/jats:styled-content> imaging by <jats:sup>89<\/jats:sup>Zr\u2010labeled scFv was done in mice bearing xenografts with <jats:styled-content style=\"fixed-case\">MSLN<\/jats:styled-content>\u2010expressing cancer cells, and tumor legions were successfully visualized. The scFv variant established in the present study may be potentially useful for cancer diagnosis by <jats:styled-content style=\"fixed-case\">PET<\/jats:styled-content> imaging.<\/jats:p>","DOI":"10.1111\/cas.14150","type":"journal-article","created":{"date-parts":[[2019,8,28]],"date-time":"2019-08-28T22:02:00Z","timestamp":1567029720000},"page":"2722-2733","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Novel single\u2010chain variant of antibody against mesothelin established by phage library"],"prefix":"10.1111","volume":"110","author":[{"given":"Hiromasa","family":"Yakushiji","sequence":"first","affiliation":[{"name":"Department of Cell Chemistry Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences Okayama Japan"},{"name":"Department of Medical Life Science Faculty of Medical Bioscience Kyushu University of Health and Welfare Miyazaki 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