title: [Preparation of antibodies against phosphorylated YB-1 and investigation of the clinical value as a biomarker to monitor hepatocellular carcinoma with pulmonary metastasis].
abstract: OBJECTIVE
  To prepare antibodies (pAbs) against phosphorylated Y-box binding protein 1 (pYB-1), perform qualitative detection of the ascites/pYB-1 ratio in patients with hepatocellular carcinoma with pulmonary metastasis (HCC-PM), and assess the clinical significance of the ascites/pYB-1 ratio as a diagnostic biomarker for HCC-PM.
  
  
  METHODS
  Bioinformatic prediction and chemical synthesis was used to identify and generate the YB-1 polypeptide with phosphorylation at serine position 102 (KYLRSVGDG). Rabbits were immunized with the YB-1 polypeptide coupled to a carrier protein. Protein A affinity chromatography was used to prepare highly-purified pAbs.ELISA and SDS-PAGE were used to determine concentration and purity of the pAbs. A total of 109 ascites specimens were collected from patients (36 cases of HCC,44 cases HCC-PM, and 29 cases of liver cirrhosis) and concentrated to obtain the pYB-1. Western blotting was used to qualitatively detect pYB-1 in ascites. Regression analysis and receiver operating characteristic (ROC) curve analysis were used to assess the qualitative data.
  
  
  RESULTS
  The prepared pAbs had a concentration of more than or equal to 1:1 * 106 and high purity. The pAbs/YB-1S102 specifically recognized endogenous pYB-1S102. The pYB-1S102 detected in ascites specimens from patients with HCC and HCC-PM, and the positive rate of detection was 30.6% and 77.3% respectively (P < 0.01).The pYB-1S102 showed sensitivity of 77.3% and a accuracy rate of 73.8% for diagnosis of HCC-PM.
  
  
  CONCLUSIONS
  Detection of pYB-1S102 in ascites could be a useful biomarker for diagnosis and metastasis monitoring in patients with HCC.
authors.affiliation: Department of Laboratory, the Second Affiliated Hospital, Chongqing Medical University, Chongqing 400010, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Nie
authors.forename: Jing
authors.initials: J
authors.lastname: Shi
authors.forename: Ding
authors.initials: D
authors.lastname: Wang
authors.forename: Li
authors.initials: L
authors.lastname: Mai
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhao
authors.forename: Qin
authors.initials: Q
authors.lastname: Hu
authors.forename: Weixian
authors.initials: W
authors.lastname: Chen
authors.forename: Pu
authors.initials: P
authors.lastname: Li
mesh_terms: D000906:Antibodies; D014408:Biomarkers, Tumor; D015153:Blotting, Western; D006528:Carcinoma, Hepatocellular; D006801:Humans; D008103:Liver Cirrhosis; D008113:Liver Neoplasms; D009362:Neoplasm Metastasis; D010766:Phosphorylation; D012372:ROC Curve
chemical_list: D000906:Antibodies; D014408:Biomarkers, Tumor