title: A novel monoclonal antibody GMab-m1 specifically recognizes IDH1-R132G mutation.
abstract: Isocitrate dehydrogenase 1 (IDH1) mutations occur in gliomas, acute myeloid leukemias, and cartilaginous tumors. While IDH1 catalyzes the oxidative carboxylation of isocitrate to α-ketoglutarate in cytosol, mutated IDH1 proteins possess the ability to change α-ketoglutarate into onco-metabolite R(-)-2-hydroxyglutarate (2-HG). To data, two monoclonal antibodies (mAbs), which are specific for IDH1 mutations have been established: clone HMab-1 against IDH1-R132H and clone SMab-1 against IDH1-R132S. However, specific mAbs against IDH1-R132G, which are useful for immunohistochemical analysis, have not been reported. To establish IDH1-R132G-specific mAbs, we immunized mice with IDH1-R132G-containing peptides. Established mAb GMab-m1 reacted with the IDH1-R132G peptide, but not with IDH1-wild type (WT) in ELISA. Western-blot analysis also showed that GMab-m1 reacted with the IDH1-R132G recombinant proteins, not with IDH1-WT or other IDH1 mutants, indicating that GMab-m1 is IDH1-R132G-specific. Furthermore, GMab-m1 specifically stained the IDH1-R132G-expressing glioma cells in immunohistochemistry. This is the first report to establish anti-IDH1-R132G-specific mAbs, which is useful in immunohistochemistry of IDH1-R132G-bearing tumors.
authors.affiliation: Regional Innovation Strategy Support Program, Tohoku University Graduate School of Medicine, 2-1 Seiryo-machi, Sendai, Miyagi 980-8575, Japan. yukinari-k@bea.hi-ho.ne.jp
authors.forename: Yukinari
authors.initials: Y
authors.lastname: Kato
authors.forename: Atsushi
authors.initials: A
authors.lastname: Natsume
authors.forename: Mika Kato
authors.initials: MK
authors.lastname: Kaneko
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D000911:Antibodies, Monoclonal; D001120:Arginine / Q000235:genetics; D002460:Cell Line; D006224:Cricetinae; D005910:Glioma / Q000175:diagnosis* / Q000201:enzymology / Q000235:genetics; D005998:Glycine / Q000235:genetics; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D007521:Isocitrate Dehydrogenase / Q000032:analysis* / Q000235:genetics* / Q000276:immunology; D051379:Mice; D009154:Mutation / Q000276:immunology; D051381:Rats; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D001120:Arginine; D007521:Isocitrate Dehydrogenase; C543588:IDH1 protein, human; D005998:Glycine