한빛사 논문
Hyunsoo Kim,1,2,3,† Areum Sohn,3,† Injoon Yeo,1 Su Jong Yu,4 Jung-Hwan Yoon,4 and Youngsoo Kim1,2,3,*
1Department of Biomedical Engineering; 2Institute of Medical and Biological Engineering, Medical Research Center; 3Department of Biomedical Sciences; 4Department of Internal Medicine and Liver Research Institute, Seoul National University College of Medicine, Seoul, Republic of Korea.
*Address correspondence to this author at: Department of Biomedical Engineering, Seoul
National University College of Medicine, 28 Yongon-Dong, Jongno-gu, Seoul 110-799,
Republic of Korea.
†H. Kim and A. Sohn contributed equally to this work.
Abstract
BACKGROUND
Lens culinaris agglutinin-reactive fraction of α-fetoprotein (AFP-L3) is a serum biomarker for hepatocellular carcinoma (HCC). AFP-L3 is typically measured by liquid-phase binding assay (LiBA). However, LiBA does not always reflect AFP-L3 concentrations because of its low analytical sensitivity. Thus, we aimed to develop an analytically sensitive multiple reaction monitoring–mass spectrometry (MRM-MS) assay to quantify AFP-L3 in serum.
METHODS
The assay entailed the addition of a stable isotope-labeled internal standard protein analog, the enrichment of AFP using a monoclonal antibody, the fractionation of AFP-L3 using L. culinaris agglutinin lectin, deglycosylation, trypsin digestion, online desalting, and MRM-MS analysis. The performance of the MRM-MS assay was compared with that of LiBA in 400 human serum samples (100 chronic hepatitis, 100 liver cirrhosis, and 200 HCC). Integrated multinational guidelines were followed to validate the assay for clinical implementation.
RESULTS
The lower limit of quantification of the MRM-MS assay (0.051 ng/mL) for AFP-L3 was less than that of LiBA (0.300 ng/mL). Thus, AFP-L3, which was not observed by LiBA in HCC samples (n = 39), was detected by the MRM-MS assay, improving the clinical value of AFP-L3 as a biomarker by switching to a more analytical sensitive platform. The method was validated, meeting all the criteria in integrated multinational guidelines.
CONCLUSIONS
Because of the lower incidence of false-negative findings, the MRM-MS assay is more suitable than LiBA for early detection of HCC.
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