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However, other novel pemphigus\u2010related autoantigens remain to be identified. In this study, <jats:bold>i<\/jats:bold>mmunoblotting for serum from an atypical autoimmune bullous disease patient identified an unknown 175 <jats:styled-content style=\"fixed-case\">kD<\/jats:styled-content>a protein. Subsequent studies using two\u2010dimensional gel electrophoresis, immunoblotting and mass\u2010spectrometry identified the 175 kDa protein as early endosome antigen 1 (<jats:styled-content style=\"fixed-case\">EEA<\/jats:styled-content>1). This finding was confirmed by subsequent immunological studies, including indirect immunofluorescence of skin and cultured keratinocytes, two\u2010dimensional gel electrophoresis and immunoblotting with anti\u2010<jats:styled-content style=\"fixed-case\">EEA<\/jats:styled-content>1 polyclonal antibody, and preabsorption with <jats:styled-content style=\"fixed-case\">EEA<\/jats:styled-content>1 recombinant protein. Finally, we developed a novel <jats:styled-content style=\"fixed-case\">BIOCHIP<\/jats:styled-content> assay using full\u2010length <jats:styled-content style=\"fixed-case\">EEA<\/jats:styled-content>1 recombinant protein to detect anti\u2010<jats:styled-content style=\"fixed-case\">EEA<\/jats:styled-content>1 antibodies. However, none of 35 sera from various types of pemphigus showed anti\u2010<jats:styled-content style=\"fixed-case\">EEA<\/jats:styled-content>1 antibodies in the <jats:styled-content style=\"fixed-case\">BIOCHIP<\/jats:styled-content> assay, with the exception of the serum from the index case. In addition, various findings in the index case did not suggest pathogenic role of anti\u2010<jats:styled-content style=\"fixed-case\">EEA<\/jats:styled-content>1 autoantibodies. Therefore, although we successfully identified the 175 <jats:styled-content style=\"fixed-case\">kD<\/jats:styled-content>a protein reacted by a serum of an atypical pemphigus\u2010like patient as <jats:styled-content style=\"fixed-case\">EEA<\/jats:styled-content>1, novel <jats:styled-content style=\"fixed-case\">BIOCHIP<\/jats:styled-content> study for other pemphigus sera indicated that <jats:styled-content style=\"fixed-case\">EEA<\/jats:styled-content>1 is not a common and pathogenic autoantigen in pemphigus.<\/jats:p>","DOI":"10.1111\/exd.12981","type":"journal-article","created":{"date-parts":[[2016,2,25]],"date-time":"2016-02-25T04:09:26Z","timestamp":1456373366000},"page":"368-374","source":"Crossref","is-referenced-by-count":0,"title":["Anti\u2010early endosome antigen 1 autoantibodies were detected in a pemphigus\u2010like patient but not in the majority of pemphigus diseases"],"prefix":"10.1111","volume":"25","author":[{"given":"Ryuhei","family":"Nishikawa","sequence":"first","affiliation":[{"name":"Department of Dermatology Kurume University School of Medicine, and Kurume University Institute of Cutaneous Cell Biology Kurume Fukuoka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hitoshi","family":"Takahashi","sequence":"additional","affiliation":[{"name":"Department of Dermatology Shimane University Faculty of Medicine Izumo Shimane Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mitsuhiro","family":"Matsuda","sequence":"additional","affiliation":[{"name":"Department of Dermatology Kurume University School of Medicine, and Kurume University Institute of Cutaneous Cell Biology Kurume Fukuoka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kaoru","family":"Imaoka","sequence":"additional","affiliation":[{"name":"Department of Dermatology Shimane University Faculty of Medicine Izumo Shimane Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masahiro","family":"Ogawa","sequence":"additional","affiliation":[{"name":"Department of Dermatology Kurume University School of Medicine, and Kurume University Institute of Cutaneous Cell Biology Kurume Fukuoka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kwesi","family":"Teye","sequence":"additional","affiliation":[{"name":"Department of Dermatology Kurume University School of Medicine, and Kurume University Institute of Cutaneous Cell Biology Kurume Fukuoka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Atsunari","family":"Tsuchisaka","sequence":"additional","affiliation":[{"name":"Department of Dermatology Kurume University School of Medicine, and Kurume University Institute of Cutaneous Cell Biology Kurume Fukuoka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hiroshi","family":"Koga","sequence":"additional","affiliation":[{"name":"Department of Dermatology Kurume University School of Medicine, and Kurume University Institute of Cutaneous Cell Biology Kurume Fukuoka Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lars","family":"Komorowski","sequence":"additional","affiliation":[{"name":"Institute for Experimental Immunology Affiliated to Euroimmun AG Luebeck Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christian","family":"Probst","sequence":"additional","affiliation":[{"name":"Institute for Experimental Immunology Affiliated to Euroimmun AG Luebeck Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takahisa","family":"Hachiya","sequence":"additional","affiliation":[{"name":"Antibody Engineering Department\/Manufacturing Division Medical &amp; Biological Laboratories Co., Ltd. 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