Wu Miao, Zhai Songhui, Gao Julia, Wei Dapeng, Xue Jianxin, Zhou Yuxi, Li Nan, Hu Lijuan
Department of Immunology, West China School of Basic Medical Sciences and Forensic Medicine, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China.
Oncol Lett. 2019 Mar;17(3):3103-3112. doi: 10.3892/ol.2019.9943. Epub 2019 Jan 17.
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 (2.6×10 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.
甘氨胆酸（GCA）是一种新发现的肝细胞癌（HCC）生物标志物。然而，由于抗GCA单克隆抗体（mAb）的可得性有限以及检测方法受限，GCA的临床病理研究尚未广泛开展。在本研究中，以与人血清白蛋白偶联的人GCA作为免疫原免疫BALB/c小鼠，制备并鉴定了一种新型抗GCA mAb。抗GCA mAb重链和轻链的亚型分别检测为IgG2a和κ，具有高亲和力常数（2.6×10 mol/l）。该抗GCA mAb以高特异性和敏感性结合GCA，50%抑制率为77.09 ng/ml。本研究还利用该抗GCA mAb建立了一种快速、灵敏且高效的间接竞争ELISA分析法，以检测不同HCC细胞系产生的GCA水平。因此，本研究可能成功开发出一种早期HCC诊断的新方法，也为HCC的进一步研究和治疗提供思路。
