biostudies-literatureUnknown7(1)Kumar ALC-MS and immunoassay can detect protein biomarkers. Immunoassays are more commonly used but can potentially be outperformed by LC-MS. These techniques have limitations including the necessity to generate separate calibration curves for each biomarker. We present a rapid mass spectrometry-based assay utilising a universal calibration curve. For the first time we analyse clinical samples using the HeavyPeptide IGNIS kit which establishes a 6-point calibration curve and determines the biomarker concentration in a single LC-MS acquisition. IGNIS was tested using apolipoprotein F (APO-F), a potential biomarker for non-alcoholic fatty liver disease (NAFLD). Human serum and IGNIS prime peptides were digested and the IGNIS assay was used to quantify APO-F in clinical samples. Digestion of IGNIS pScientific reports12072https://www.ebi.ac.uk/biostudies/studies/S-EPMC5608892biostudies-literatureAbsolute quantitation of disease protein biomarkers in a single LC-MS acquisition using apolipoprotein F as an example.PMC5608892Cobbold JZitzmann NThursz MKumar AGangadharan BfalseAbsolute quantitation of disease protein biomarkers in a single LC-MS acquisition using apolipoprotein F as an example.LC-MS and immunoassay can detect protein biomarkers. Immunoassays are more commonly used but can potentially be outperformed by LC-MS. These techniques have limitations including the necessity to generate separate calibration curves for each biomarker. We present a rapid mass spectrometry-based assay utilising a universal calibration curve. For the first time we analyse clinical samples using the HeavyPeptide IGNIS kit which establishes a 6-point calibration curve and determines the biomarker concentration in a single LC-MS acquisition. IGNIS was tested using apolipoprotein F (APO-F), a potential biomarker for non-alcoholic fatty liver disease (NAFLD). Human serum and IGNIS prime peptides were digested and the IGNIS assay was used to quantify APO-F in clinical samples. Digestion of IGNIS p2017-01-01T00:00:00Z2017 Sep2026-05-06T06:32:42.094Z2019-03-27T02:57:04ZS-EPMC56088922893589510.1038/s41598-017-12229-2