Functional expression, characterization, and application of human S100B

被引:7
作者
Wu, Lei [1 ]
Zhou, Xuesi [1 ]
Xiao, Zhigang [1 ]
Gao, Xiujie [1 ]
Liu, Ziquan [1 ,2 ,3 ]
Zhang, Zhiqing [1 ]
Wang, Kun [1 ,3 ]
Zhu, Yulin [1 ]
Ren, Hanlin [1 ]
Wang, Tianhui [1 ]
机构
[1] Tianjin Inst Hlth & Environm Med, Performance Med Lab, 1 Dali Rd, Tianjin 300050, Peoples R China
[2] Tianjin Univ Technol, Sch Mat Sci & Engn, Tianjin 300384, Peoples R China
[3] Univ Chinese Peoples Armed Police Force, Inst Disaster Med & Publ Hlth, Affiliated Hosp Logist, Tianjin 300162, Peoples R China
基金
中国国家自然科学基金;
关键词
application; human S100B protein; monoclonal antibody; purification; MONOCLONAL-ANTIBODIES; PROTEIN; CANCER; MARKER;
D O I
10.3892/or.2017.5922
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
The EF-hand calcium-binding protein S100B presents a wide range of biological activities and functions. This binding protein is involved in various human diseases, including cancer, brain trauma and ischemia, neuro-degenerative disease (Alzheimer's disease), and psychiatric disorders. In this study, we prepared human S100B protein and its monoclonal antibodies. Human S100B protein was expressed in Escherichia coli, successfully purified by diethylaminoethyl cellulose anion-exchange chromatography, and then identified by western blot analysis. Monoclonal antibodies (mAbs) were produced by the standard hybridoma method and validated by enzyme-linked immunosorbent assay and western blot analysis. The prepared human S100B protein and its mAbs demonstrated potential biological activities. The KD of one mAb is approximately 4.72x10(-8) mol/l, and its cross reactivity is low with human S100A4, mouse S100A4, and human S100A1. Recombinant Soluble S100B can promote the migration and invasion of HeLa cells. The expression of S100B protein in tumor tissues can be detected effectively by using the prepared monoclonal antibodies. Increasing concentration of the antihuman S100B mAbs showed a reduced expression of the S100B protein. Subsequently, the expression of p53 increased significantly (P<0.05) in A375 cells. A significant increase in apoptosis in A375 cells was observed with increasing S100B mAb concentration. Results showed that our prepared S100B mAbs were suitable for detecting S100B expression in human tissues, furnishing promising tools for further functional investigation and clinical applications.
引用
收藏
页码:2309 / 2316
页数:8
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