Technologies

Solutions

Researchers from DKFZ and University Hospital of Heidelberg developed a test for detecting D2HG in diverse samples by measuring the production of the reduced state of the dye. The technique can be used for diagnosis but in addition monitoring a D2HG-associated disease of a patient.

Figure

Figure 1: D2HG was diluted in water, blood serum, and urine to get a standard curve (0-375 pmol = 0-15 µM). Samples were prepared according to the protocol in Figure 2 and assayed with the diaphorase/ resazurin read-out. In the graph, the relative fluorescence (RFU) is blotted against the D2HG concentration

Figure 2: Measurement of sera of patients suffering from acute myeloid leukemia (AML). These patients carry an IDH1 mutation which results in the production of D2HG. Comparison between gas chromatography-mass spectrometry (GC-MS) (single determination) and enzyme assay of the invention using the diaphorase/ resazurin read-out (in triplicate). In comparison, sera of healthy persons who did not carry an IDH1 mutation have been tested. These samples were D2HG negative.

References

1. “Detection of 2-hydroxyglutarate in formalin-fixed paraffin-embedded glioma specimens by gas chromatography/mass spectrometry.” in Brain Pathol. 2012 Jan;22(1):26-31 by Sahm F et al.. See at: www.ncbi.nlm.nih.gov/pubmed/21631627
2. “Enzymatic assay for quantitative analysis of (D)- 2-hydroxyglutarate.” in Acta Neuropathol. 2012 Dec;124(6):883-91 by Balss J et al.. See at: www.ncbi.nlm.nih.gov/pubmed/23117877
3. “Pan‑ mutant IDH1 inhibitor BAY 1436032 for effective treatment of IDH1 mutant astrocytoma in vivo.” In Acta Neuropathol. 2017 Jan 25. PMID: 28124097 by Stefan Pusch et al.. See at: www.ncbi.nlm.nih.gov/pubmed/
4. “Pan-mutant-IDH1 inhibitor BAY1436032 is highly effective against human IDH1 mutant acute myeloid leukemia in vivo.” In Leukemia, Jan 2017; doi: 10.1038/leu.2017.46. by A Chaturvedi et al..