Glycocholic acid (GCA) is a novel identified biomarker for hepatocellular carcinoma (HCC). However, clinical pathological study of GCA has not been extensive due to the limited availability of anti-GCA monoclonal antibodies (mAbs) and restricted detection methods. In the present study, using human GCA conjugated with bovine serum albumin as the immunogen to immunize BALB/c mice, a novel anti-GCA mAb was generated and characterized. The isotypes of heavy chain and light chain of anti-GCA mAb were examined to be IgG2a and κ, respectively, with a high affinity constant (2.6×108 mol/l). The anti-GCA mAb binds GCA with high specificity and sensitivity, and the 50% inhibitory rate was 77.09 ng/ml. The present study also established a rapid, sensitive and efficient indirect competitive ELISA analysis using this anti-GCA mAb to detect the level of GCA produced by different HCC cell lines. Therefore, the present study may successfully develop a novel method for early HCC diagnosis, and also provide insights for further research and treatment of HCC.
Keywords: glycocholic acid; hepatocellular carcinoma; indirect competitive enzyme-linked immunoassay; monoclonal antibody.