Establishment of a Multi-Specific Monoclonal Antibody MsMab-1 Recognizing Both IDH1 and IDH2 Mutations

摘要: Mutations of isocitrate dehydrogenase 1 (IDH1) and 2 (IDH2) have been reported in gliomas, cartilaginous tumors, acute myeloid leukemias. IDH mutations are specific to a single codon the conserved functionally important arginine 132 residue (R132) IDH1 or 172 (R172) IDH2 gliomas. Although catalyze oxidative carboxylation α-ketoglutarate cytosol mitochondria, respectively, mutated IDH1/2 proteins can possess ability change an oncometabolite R(-)-2-hydroxyglutarate. We established several monoclonal antibodies (mAbs) for mutations. However, no multi-specific mAb against has reported. For this study, we immunized mice with IDH1-R132G peptide 19 amino acids (GGVKPIIIGGHAYGDQYRA), novel MsMab-1 that recognizes IDH1-R132G, but not wild type enzyme-linked immunosorbent assay (ELISA). It is particularly interesting all mutants (R132H, R132C, R132S, R132G, R132L) ELISA. Western blot analysis also revealed reacted recombinant IDH1-R132H, IDH1-R132S, other mutations, indicating anti-mutated mAb. Unexpectedly, IDH2-R172M protein, despite shows only 73.7% identity equivalent portion (GGTKPITIGMHAHGDQYKA). Moreover, stained IDH1-R132S IDH1-R132G-expressing glioma cells immunohistochemistry. This report first establish mAb, expected be useful immunohistochemical determination mutation-bearing tumors.