Supplementary Table 1:

Estimated assay validation parameters including limit of detection (LOD), limit of quantification (LOQ) and Z´-factor for different sample origins up to 50 µM D2HG per well. Values of LOD and LOQ are given in µM D2HG. Z´-factor is dimensionless value.

Sample origin / LOD / LOQ / Z´-factor
ddH20 / 0.13 / 0.44 / 0.95
Cell culture medium (+phenolred) / 0.74 / 2.47 / 0.9
Cell culture medium
(-phenolred) / 0.53 / 1.77 / 0.91
Blood serum / 0.83 / 2.77 / 0.85
Urine / 27.4 / 91.8 / 0.11


Supplementary Table 2:

Overview of patients screened for D2HG with information about diagnosis and IDH1/2 mutation status (sequenced residues: IDH1 R100 and R132; IDH2 R140 and R172)

Patient / Diagnosis / IDH 1/2 mutation status / D2HG screen of
P1 / secGBM / IDH1 R132G / frozen tissue, paraffin
P2 / AIII / IDH1 R132H / frozen tissue, paraffin
P3 / OII / IDH1 R132H / frozen tissue, paraffin
P4 / OAII / IDH1 R132H / frozen tissue, paraffin
P5 / AIII / IDH1 R132H / frozen tissue, paraffin
P6 / AII / IDH1 R132H / frozen tissue
P7 / pGBM / wt / frozen tissue, paraffin
P8 / pGBM / wt / frozen tissue, paraffin
P9 / pGBM / wt / frozen tissue, paraffin
P10 / pGBM / wt / frozen tissue, paraffin
P11 / pGBM / wt / frozen tissue, paraffin
P12 / pGBM / wt / frozen tissue, paraffin
P13 / pGBM / wt / frozen tissue
P14 / pGBM / wt / frozen tissue
P15 / pGBM / wt / frozen tissue
P1-AML / AML / IDH1 R132H / serum
P2-AML / AML / IDH1 R132H / serum
P3-AML / AML / IDH1 R132C / serum
P4-AML / AML / IDH2 R140Q / serum
P5-AML / AML / IDH2 R140Q / serum
P6-AML / AML / wt / serum
P7-AML / AML / wt / serum
P8-AML / AML / wt / serum
P9-AML / AML / wt / serum
P10-AML / AML / wt / serum
P11-AML / AML / wt / serum
P12-AML / AML / wt / serum
P13-AML / AML / wt / serum


Supplementary Table 3:

Intrarun (n=5) and Interrun (n=5) measurements with 0.5, 5 and 50 µM D2HG solved in medium and serum

D2HG [µM] / Medium / Serum
added / found
Intrarun / Interrun / Intrarun / Interrun
0.5 µM D2HG / 0.55 (0.14) / 0.56 (0.2) / 1.28 (0.34) / 1.21 (0.17)
5 µM D2HG / 5.52 (0.25) / 4.95 (0.94) / 5.22 (0.52) / 5.44 (0.30)
50 µM D2HG / 50.4 (1) / 51.4 (1.62) / 48 (0.75) / 50.3 (2.23)
Accuracy [%] / Medium / Serum
Intrarun / Interrun / Intrarun / Interrun
0.5 µM D2HG / 9.16 / 11.5 / 156.3 / 141.5
5 µM D2HG / 10.5 / -1.01 / 4.49 / 8.72
50 µM D2HG / 0.77 / 2.73 / -3.98 / 0.54