title: Evaluation of a magnetic particles-based chemiluminescence enzyme immunoassay for Golgi protein 73 in human serum.
abstract: BACKGROUND
  Golgi protein 73 (GP73) is regarded as a potential serum biomarker for early diagnosis of hepatocellular carcinoma (HCC). We developed a rapid magnetic particles-based chemiluminescence enzyme immunoassay (MPs-CLEIA) for the determination of serum GP73.
  
  
  METHODS
  Fluorescein isothiocyanate (FITC) and alkaline phosphatase (ALP) were used to label 2 different monoclonal antibodies to GP73. Serum GP73 was captured with labeled antibodies and formed a sandwiched immunoreaction. The magnetic particles (MPs) coated with anti-FITC antibody were used as a means of separation of the GP73 protein from other serum proteins. After adding the enzyme substrate solution, the relative light unit (RLU) was measured. A MPs-CLEIA for serum GP73 was established and evaluated.
  
  
  RESULTS
  The RLU was directly proportional to the concentration of GP73. The method linearity was 5-600 μg/l. Limit of the blank was 2.19 μg/l. The intra- and inter-assay imprecision was <3% and <5%, respectively. The average recoveries were between 95% and 105%. The proposed method showed a good correlation with a commercial ELISA assay (r=0.983, p<0.001). We also evaluated the efficiency of serum GP73 measurement for the diagnosis of HCC using this assay. The area under the receiver operating characteristic curve was 0.822 (95% CI, 0.73-0.89), and the sensitivity and specificity, with cut-off value of 115.6 μg/l, were 75.4% and 92.1%, respectively.
  
  
  CONCLUSIONS
  The proposed method demonstrates an acceptable performance for quantifying serum GP73. This assay could be appropriate for routine use in clinical laboratories.
authors: Department of Medical Laboratory, Beijing Tongren Hospital, Capital Medical University, Beijing, PR China.
authors: Xiangyi
authors: X
authors: Liu
authors: Department of Medical Laboratory, Beijing Tongren Hospital, Capital Medical University, Beijing, PR China.
authors: Xiaohua
authors: X
authors: Wan
authors: Beijing Ranos Medical Technology Co., Ltd, Beijing, PR China.
authors: Sheng
authors: S
authors: Lu
authors: Department of Surgery, Beijing Tongren Hospital, Capital Medical University, Beijing, PR China.
authors: Lijun
authors: L
authors: Zhang
authors: Beijing Hotgen Biotechnology Co., Ltd., Beijing, PR China.
authors: Shaohua
authors: S
authors: Yu
authors: Department of Medical Laboratory, Beijing Tongren Hospital, Capital Medical University, Beijing, PR China. Electronic address: 2013luxinxin@sina.com.
authors: Xinxin
authors: X
authors: Lu
mesh_terms: D000469:Alkaline Phosphatase / Q000737:chemistry; D000911:Antibodies, Monoclonal / Q000737:chemistry; D015415:Biomarkers / Q000097:blood; D002138:Calibration; D006528:Carcinoma, Hepatocellular / Q000097:blood / Q000175:diagnosis* / Q000235:genetics; D016022:Case-Control Studies; D004797:Enzyme-Linked Immunosorbent Assay; D005234:Fatty Liver / Q000097:blood / Q000175:diagnosis* / Q000235:genetics; D005260:Female; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D015870:Gene Expression; D006509:Hepatitis B / Q000097:blood / Q000175:diagnosis* / Q000235:genetics; D006801:Humans; D008113:Liver Neoplasms / Q000097:blood / Q000175:diagnosis* / Q000235:genetics; D059346:Magnets; D008297:Male; D008565:Membrane Proteins / Q000097:blood* / Q000235:genetics; D008875:Middle Aged; D009367:Neoplasm Staging; D012372:ROC Curve; D015203:Reproducibility of Results; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D015415:Biomarkers; C410824:GOLM1 protein, human; D008565:Membrane Proteins; D000469:Alkaline Phosphatase; D016650:Fluorescein-5-isothiocyanate
keywords: Chemiluminescence enzyme immunoassay; Evaluation; Golgi protein 73; Hepatocellular carcinoma; Magnetic particles