Development of a quantitative serum assay of Golgi protein 73 in hepatocellular carcinoma using xMAP technology
10.3760/cma.j.issn.0253-3766.2019.05.006
- VernacularTitle: 应用液态悬浮芯片技术建立肝癌血清标志物蛋白GP73的检测方法
- Author:
Yun WU
1
;
Yipeng WANG
2
;
Jie MA
3
;
Yonghong ZHANG
1
;
Huanqin SUN
1
;
Jianping SUN
1
;
Zikang WANG
1
;
Jie XU
1
;
Yanchao DAI
1
;
Ning LI
4
Author Information
1. Department of Biomedical Information Center, Beijing You′an Hospital, Capital Medical University, Beijing 100069, China
2. Department of Prenatal Diagnosis Center, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100026, China
3. Beijing Qingyuan Bio-Technologies Inc, Beijing, 100102, China
4. Department of Surgery, Beijing You′an Hospital, Capital Medical University, Beijing 100069, China
- Publication Type:Clinical Trail
- Keywords:
Hepatocellular carcinoma;
Golgi protein 73;
Antibody screening;
Liquid chip
- From:
Chinese Journal of Oncology
2019;41(5):351-356
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish a quantitative assay of serum Golgi protein 73 (GP73) using xMAP technology and evaluate its performance.
Methods:Monoclonal antibodies against GP73 were prepared and purified, and antibody pair screening was performed by double-antibody sandwich enzyme-linked immunosorbent assay. The screened antibodies were used to construct a Luminex liquid chip detection system, and the analysis performance of the detection system was evaluated. The serum levels of GP73 were detected in 90 clinical samples from healthy controls and patients with chronic hepatitis B infection (CHB) and hepatocellular carcinoma (HCC).
Results:Five anti-GP73 monoclonal antibodies were prepared and purified, and 5 antibody pairs were successfully screened. The Luminex liquid chip detection system of GP73 was successfully constructed using 8F10D1 and 10B9F11 antibody pairs. The analytical performance evaluation showed that the sensitivity of this system was 0.25 ng/ml and the dynamic range was 0.25-100 ng/ml. No cross reactivity was observed. The intra- and inter-assay variation for GP73 was <8% and <11%, respectively. The recovery was 83%-92%. The linear regression equation was y=1.141x+ 6.436 (r2=0.998 4, P<0.001). The GP73 concentrations in the serum samples of healthy control, CHB group, and HCC group were 42.8 (38.68, 55.90) ng/ml, 61.49 (43.59, 81) ng/ml, and 122.78 (49.36 liter, 264.55) ng/ml, respectively. The levels of GP73 in HCC group were significantly higher than those in CHB group and healthy controls (P<0.05). Moreover, the levels of GP73 in CHB group were significantly higher than those in healthy controls (P<0.05).
Conclusions:A liquid chip detection system of GP73 was successfully constructed. It provides a powerful tool for the clinical application of GP73 in the future.
