title: Establishment of a novel monoclonal antibody SMab-1 specific for IDH1-R132S mutation.
abstract: Isocitrate dehydrogenase 1 (IDH1) mutations, which are early and frequent genetic alterations in gliomas, are specific to a single codon in the conserved and functionally important Arginine 132 (R132) in IDH1. We earlier established a monoclonal antibody (mAb), IMab-1, which is specific for R132H-containing IDH1 (IDH1-R132H), the most frequent IDH1 mutation in gliomas. To establish IDH1-R132S-specific mAb, we immunized mice with R132S-containing IDH1 (IDH1-R132S) peptide. After cell fusion using Sendai virus envelope, IDH1-R132S-specific mAbs were screened in ELISA. One mAb, SMab-1, reacted with the IDH1-R132S peptide, but not with other IDH1 mutants. Western-blot analysis showed that SMab-1 reacted only with the IDH1-R132S protein, not with IDH1-WT protein or IDH1 mutants, indicating that SMab-1 is IDH1-R132S-specific. Furthermore, SMab-1 specifically stained the IDH1-R132S-expressing glioblastoma cells in immunocytochemistry and immunohistochemistry, but did not react with IDH1-WT or IDH1-R132H-containing glioblastoma cells. We newly established an anti-IDH1-R132S-specific mAb SMab-1 for use in diagnosis of mutation-bearing gliomas.
authors.affiliation: Molecular Tumor Marker Research Team, The Oncology Research Center, Advanced Molecular Epidemiology Research Institute, Yamagata University Faculty of Medicine, 2-2-2 Iida-nishi, Yamagata 990-9585, Japan.
authors.forename: Mika Kato
authors.initials: MK
authors.lastname: Kaneko
authors.forename: Wei
authors.initials: W
authors.lastname: Tian
authors.forename: Shingo
authors.initials: S
authors.lastname: Takano
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Suzuki
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Sawa
authors.forename: Yasukazu
authors.initials: Y
authors.lastname: Hozumi
authors.forename: Kaoru
authors.initials: K
authors.lastname: Goto
authors.forename: Kentaro
authors.initials: K
authors.lastname: Yamazaki
authors.forename: Chifumi
authors.initials: C
authors.lastname: Kitanaka
authors.forename: Yukinari
authors.initials: Y
authors.lastname: Kato
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology*; D045744:Cell Line, Tumor; D005910:Glioma / Q000175:diagnosis* / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007521:Isocitrate Dehydrogenase / Q000032:analysis* / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C
chemical_list: D000911:Antibodies, Monoclonal; D007521:Isocitrate Dehydrogenase; C543588:IDH1 protein, human