Metidq version 5 4 8 db100 boron 2607

Targeted metabolomics on cell-free ascites and serum of a subset of 19 HGSC patients was performed using AbsoluteIDQ p180 kits (Biocrates Life Sciences AG, Innsbruck, Austria). The kit allows the identification and (semi-) quantification of metabolites by LC- and flow injection analysis (FIA)-MRM. The samples were analyzed on an AB SCIEX QTrap 4000 mass spectrometer (Framingham, MA, USA) using an Agilent 1200 RR HPLC system (Agilent Technologies, Santa Clara, CA, USA), which were operated with Analyst 1.6.2 (AB SCIEX). The chromatographic column was obtained from Biocrates. The serum samples and additional blanks, calibration standards and quality controls were prepared according to the user manual. All amino acids and biogenic amines were derivatized with phenylisothiocyanate. The experiments were validated using the supplied software (MetIDQ, Version 5-4-8-DB100-Boron-2607, Biocrates). Histamine, methioninesulfoxide, symmetric dimethylarginine, α-aminoadipic acid and trans-4-hydroxyproline did not pass quality controls and therefore, were not considered for data interpretation.

Targeted metabolomics in serum was performed using AbsoluteIDQ p180 kits (Biocrates Life Sciences AG, Innsbruck, Austria)20 (link). The kit allows the identification and (semi-) quantification of metabolites by liquid chromatography (LC)- and flow injection analysis (FIA)-multiple reaction monitoring (MRM). The samples were analyzed on an AB SCIEX QTrap 4000 mass spectrometer (Framingham, MA, USA) using an Agilent 1200 RR HPLC system (Agilent Technologies, Santa Clara, CA, USA), which were operated with Analyst 1.6.2 (AB SCIEX). The chromatographic column was obtained from Biocrates. The serum samples and additional blanks, calibration standards and quality controls were prepared according to the user manual. All amino acids and biogenic amines were derivatized with phenylisothiocyanate. The experiments were validated using the supplied software (MetIDQ, Version 5-4-8-DB100-Boron-2607, Biocrates). Metabolite concentrations were compared to PD-L1 abundances as described in the Luminex analyses section. A false discovery rate (FDR) cut-off of 5% was used.

Targeted metabolomics was performed using the AbsoluteIDQ p180 kit (Biocrates Life Sciences AG, Innsbruck, Austria). The kit allows the identification and quantitation of metabolites by LC- and flow injection analysis (FIA)-MRM. All serum samples were stored at −80 °C until being analyzed on an AB SCIEX QTrap 4000 mass spectrometer using an Agilent 1200 RR HPLC system, which were operated with Analyst 1.6.2 (AB SCIEX). The chromatographic column was obtained from Biocrates. The serum samples and additional blanks, calibration standards and quality controls were prepared according to the user manual. The experiments were validated with the supplied software (MetIDQ, Version 5-4-8-DB100-Boron-2607, Biocrates Life Sciences, Innsbruck, Austria). Quality controls were analyzed every 20^th sample. Data was analyzed and visualized with R Studio. For metabolites that were below the limit of detection (LOD), the LOD was used for calculating fold changes and p-values.