Development of a Chemiluminescence Immunoassay for Serum YB-1 and its Clinical Application as a Potential Diagnostic Marker for Hepatocellular Carcinoma

Y-box binding protein 1 (YB-1) overexpression has been shown in various tumor cells including hepatocellular carcinoma (HCC); moreover، this protein can be actively secreted.. The aim of this study was to establish a method to quantify serum YB-1 and evaluate its clinical application in the clinical diagnosis of HCC.. Patients and Recombinant YB-1 and two populations of its antibodies were prepared. A monoclonal antibody was specific to the N-terminus of YB-1 amino acids 134-160; and another was a polyclonal antibody. A sandwich-type chemiluminescence immunoassay (CLIA) was developed and evaluated. Levels of YB-1 and alpha fetoprotein (AFP) in serum samples from 105 HCC patients، 25 hepatitis B virus patients، 25 cirrhosis patients، and 50 healthy donors were detected using the established method and an AFP electrochemiluminescence kit.. The developed method was linear to 150 μg/L of YB-1 with a minimum detection limit of 0. 01 μg/L. The average recoveries were between 93. 9% and 109. 0%. The mean intra- and inter-assay coefficients of variation (CVs) were 4. 0-4. 8% and 8. 2-10. 2%، respectively. The relationship between the concentration of diluted YB-1 and the dilution ratios gave a good linear correlation coefficient of 0. 9986. The YB-1 concentration was increased in serum of HCC patients (33. 0 ± 23. 39 μg/L) compared to healthy individuals (13. 2 ± 5. 29 μg/L، P < 0. 0001)، patients with HBV (17. 9 ± 7. 49 μg/L، P = 0. 0003)، and patients with HBV cirrhosis (20. 7 ± 8. 75 μg/L، P < 0. 05). Moreover، the combination of YB-1 and alpha-fetoprotein had a high sensitivity (89. 5%) and reasonable specificity (62. 0%) in identifying HCC.. The established method has an acceptable performance in quantifying YB-1. In addition، serum YB-1 may aid in the diagnosis of HCC..