title: Establishment of novel monoclonal antibodies KMab-1 and MMab-1 specific for IDH2 mutations.
abstract: Isocitrate dehydrogenase 1/2 (IDH1/2) mutations have been detected in gliomas, cartilaginous tumors, and leukemias. IDH1/2 mutations are early and frequent genetic alterations, are specific to a single codon in the conserved and functionally important Arginine 132 (R132) in IDH1 and Arginine 172 (R172) in IDH2. We previously established several monoclonal antibodies (mAbs), which are specific for IDH1 mutations: clones IMab-1 or HMab-1 against IDH1-R132H or clone SMab-1 against IDH1-R132S. However, specific mAbs against IDH2 mutations have not been reported. To establish IDH2-mutation-specific mAbs, we immunized mice or rats with each mutation-containing IDH2 peptides including IDH2-R172K and IDH2-R172M. After cell fusion, IDH2 mutation-specific mAbs were screened in Enzyme-Linked Immunosorbent Assay (ELISA). Established mAbs KMab-1 and MMab-1 reacted with the IDH2-R172K and IDH2-R172M peptides, respectively, but not with IDH2-wild type (WT) in ELISA. Western-blot analysis also showed that KMab-1 and MMab-1 reacted with the IDH2-R172K and IDH2-R172M recombinant proteins, respectively, not with IDH2-WT or other IDH2 mutants, indicating that KMab-1 and MMab-1 are IDH2-mutation-specific. Furthermore, MMab-1 specifically stained the IDH2-R172M-expressing cells in immunocytochemistry, but did not stain IDH2-WT and other IDH2-mutation-containing cells. In immunohistochemical analysis, MMab-1 specifically stained IDH2-R172M-expressing glioma. This is the first report to establish anti-IDH2-mutation-specific mAbs, which could be useful in diagnosis of mutation-bearing tumors.
authors: Regional Innovation Strategy Support Program, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Aoba-ku, Sendai, Miyagi 980-8575, Japan.
authors: Mika Kato
authors: MK
authors: Kaneko
authors: Shunpei
authors: S
authors: Morita
authors: Yuta
authors: Y
authors: Tsujimoto
authors: Ryo
authors: R
authors: Yanagiya
authors: Kana
authors: K
authors: Nasu
authors: Hiroko
authors: H
authors: Sasaki
authors: Yasukazu
authors: Y
authors: Hozumi
authors: Kaoru
authors: K
authors: Goto
authors: Atsushi
authors: A
authors: Natsume
authors: Mika
authors: M
authors: Watanabe
authors: Toshihiro
authors: T
authors: Kumabe
authors: Shingo
authors: S
authors: Takano
authors: Yukinari
authors: Y
authors: Kato
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D016466:CHO Cells; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D005910:Glioma / Q000175:diagnosis* / Q000235:genetics*; D007521:Isocitrate Dehydrogenase / Q000032:analysis* / Q000235:genetics / Q000276:immunology*; D051379:Mice; D009154:Mutation; D051381:Rats
chemical_list: D000911:Antibodies, Monoclonal; C538784:IDH2 protein, human; D007521:Isocitrate Dehydrogenase