Establishment of a novel monoclonal antibody SMab-1 specific for IDH1-R132S mutation

Biochem Biophys Res Commun. 2011 Mar 25;406(4):608-13. doi: 10.1016/j.bbrc.2011.02.102. Epub 2011 Feb 23.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal / genetics
  • Antibodies, Monoclonal / immunology*
  • Cell Line, Tumor
  • Glioma / diagnosis*
  • Glioma / genetics
  • Humans
  • Immunohistochemistry / methods
  • Isocitrate Dehydrogenase / analysis*
  • Isocitrate Dehydrogenase / genetics*
  • Isocitrate Dehydrogenase / immunology
  • Mice
  • Mice, Inbred BALB C

Substances

  • Antibodies, Monoclonal
  • Isocitrate Dehydrogenase
  • IDH1 protein, human