A quantitative analytical method for PIVKA-II using multiple reaction monitoring-mass spectrometry for early diagnosis of hepatocellular carcinoma.
Prothrombin induced by vitamin K absence-II (PIVKA-II) is an effective tumor marker for hepatocellular carcinoma (HCC). We have developed a novel targeted mass spectrometric (MS) assay for quantifying PIVKA-II in human serum. The ideal signature peptide was selected to measure PIVKA-II concentrations on a triple quadrupole (QqQ) mass spectrometer, and the chromatography gradient was optimized for the peptide separation to minimize elution interference. Using multiple reaction monitoring-mass spectrometry (MRM-MS), good linearity (R2= 0.9988) was obtained for PIVKA-II over a range of 3 orders. We achieved a limit of detection (LOD) of 0.45 nM (31.72 ng/mL), a limit of quantification (LOQ) of 0.93 nM (65.31 ng/mL), a lower limit of quantification (LLOQ) of 0.49 nM (34.32 ng/mL), and an upper limit of quantification (ULOQ) of 1000.00 nM (70,037.00 ng/mL). The intra-day and inter-day precisions were within ±14.96%, and the accuracy ranged from 87.66 to 114.29% for QC samples at four concentrations. Compared with an established immunoassay, the correlation (R = 0.8335) was good for the measurements of PIVKA-II concentrations. This method was successfully applied to the analysis of clinical samples for normal control (n = 50), chronic hepatitis (n = 50), liver cirrhosis (n = 50), HCC (n = 50), and recovery (n = 50) serum. Graphical Abstract MRM-MS assay development for determining concentration of PIVKA-II in serum and a comparison between MRM-MS assay and immunoassay with high correlation.
Department of Biomedical Sciences, Seoul National University College of Medicine, Yongon-Dong, Seoul, 110-799, Korea.|Department of Biomedical Engineering, Seoul National University College of Medicine, Yongon-Dong, Seoul, 110-799, Korea.
Areum
A
Sohn
Department of Biomedical Sciences, Seoul National University College of Medicine, Yongon-Dong, Seoul, 110-799, Korea.|Department of Biomedical Engineering, Seoul National University College of Medicine, Yongon-Dong, Seoul, 110-799, Korea.|Institute of Medical and Biological Engineering, Medical Research Center, Seoul National University College of Medicine, Yongon-Dong, Seoul, 110-799, Korea.
Hyunsoo
H
Kim
Department of Internal Medicine, Seoul National University College of Medicine, Yongon-Dong, Seoul, 110-799, Korea.|Liver Research Institute, Seoul National University College of Medicine, Yongon-Dong, Seoul, 110-799, Korea.
Su Jong
SJ
Yu
Department of Internal Medicine, Seoul National University College of Medicine, Yongon-Dong, Seoul, 110-799, Korea.|Liver Research Institute, Seoul National University College of Medicine, Yongon-Dong, Seoul, 110-799, Korea.
Jung-Hwan
JH
Yoon
Department of Biomedical Sciences, Seoul National University College of Medicine, Yongon-Dong, Seoul, 110-799, Korea. biolab@snu.ac.kr.|Department of Biomedical Engineering, Seoul National University College of Medicine, Yongon-Dong, Seoul, 110-799, Korea. biolab@snu.ac.kr.|Institute of Medical and Biological Engineering, Medical Research Center, Seoul National University College of Medicine, Yongon-Dong, Seoul, 110-799, Korea. biolab@snu.ac.kr.
Youngsoo
Y
Kim
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D015415:Biomarkers / Q000097:blood*; D014408:Biomarkers, Tumor / Q000097:blood*; D001774:Blood Chemical Analysis / Q000379:methods; D006528:Carcinoma, Hepatocellular / Q000097:blood* / Q000175:diagnosis*; D002853:Chromatography, Liquid / Q000379:methods; D055088:Early Detection of Cancer / Q000379:methods; D005260:Female; D006801:Humans; D008113:Liver Neoplasms / Q000097:blood* / Q000175:diagnosis*; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D008875:Middle Aged; D011498:Protein Precursors / Q000097:blood*; D011516:Prothrombin; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D015415:Biomarkers; D014408:Biomarkers, Tumor; D011498:Protein Precursors; C008208:acarboxyprothrombin; D011516:Prothrombin
Absolute quantification; Hepatocellular carcinoma (HCC); Mass spectrometry; Multiple Reaction Monitoring (MRM); Prothrombin induced by vitamin K absence-II (PIVKA-II)