title: A Chemiluminescent Protein Microarray Method for Determining the Seroglycoid Fucosylation Index.
abstract: The Lens culinaris agglutinin-reactive fraction of AFP (AFP-L3) is widely used to screen for hepatocellular carcinoma (HCC) in Japan and China. We developed a chemiluminescent protein microarray for determining the AFP-L3/AFP index (the ratio of AFP-L3 to total AFP, AFP-L3%) by fixing AFP-specific antibodies and Lens culinaris lectin on aldehyde-coated glass slides. Serum samples were tested for AFP using an enzyme-linked immunosorbent assay (ELISA) to validate the microarray. AFP-L3 was detected using Hotgen Biotech glycosyl capture spin column pretreatment technology and ELISA. When the AFP cut-off value was set to 20 ng/ml, the protein microarray displayed 89.74% sensitivity and 100% specificity for HCC diagnosis, and the ELISA displayed 87.17% sensitivity and 100% specificity. When the AFP-L3% cut-off value was set to 0.1, the protein microarray displayed 56.41% sensitivity and 100% specificity for HCC diagnosis, and the ELISA displayed 53.84% sensitivity and 100% specificity. The ROC curve for the HCC diagnosis showed that the AFP area under the ROC curve (AUC = 0.996; 95% CI: 0.986-1.005) was much higher than that of AFP-L3 (AUC = 0.857; 95% CI: 0.769-0.94) and AFP-L3% (AUC = 0.827; CI: 0.730-0.924). The microarray assay used in this study is a highly sensitive, accurate, and efficient assay for the determination of the AFP-L3%.
authors.affiliation: Beijing Institute of Hepatology, Beijing YouAn Hospital, Capital Medical University, Beijing 100069, China.
authors.forename: Aiying
authors.initials: A
authors.lastname: Zhang
authors.affiliation: Sino-Swed Molecular Bio-Medicine Research Institute, Shenzhen 518057, China.
authors.forename: Sven
authors.initials: S
authors.lastname: Skog
authors.affiliation: Department of Biotechnology, Beijing Institute of Radiation Medicine, Beijing 100850, China.
authors.forename: Shengqi
authors.initials: S
authors.lastname: Wang
authors.affiliation: Peking University Health Science Center, Beijing 100191, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Ke
authors.affiliation: Beijing YouAn Hospital, Capital Medical University, Beijing 100069, China.
authors.forename: Yonghong
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Beijing YouAn Hospital, Capital Medical University, Beijing 100069, China.
authors.forename: Kang
authors.initials: K
authors.lastname: Li
authors.affiliation: Sino-Swed Molecular Bio-Medicine Research Institute, Shenzhen 518057, China.
authors.forename: Ellen
authors.initials: E
authors.lastname: He
authors.affiliation: Beijing Institute of Hepatology, Beijing YouAn Hospital, Capital Medical University, Beijing 100069, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Li
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000097:blood; D016022:Case-Control Studies; D004797:Enzyme-Linked Immunosorbent Assay; D005643:Fucose / Q000378:metabolism*; D006801:Humans; D057230:Limit of Detection; D008113:Liver Neoplasms / Q000097:blood; D049449:Luminescence; D040081:Protein Array Analysis / Q000379:methods*; D000509:alpha-Fetoproteins / Q000378:metabolism
chemical_list: D000509:alpha-Fetoproteins; D005643:Fucose