The Study of Hepatitis B Virus Detection

• Main Authors: Li, Chen-Huan, 李晨歡
• Other Authors: Li,Yaw-Kuen
• Format: Others
• Language: zh-TW
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/34dv35

碩士 === 國立交通大學 === 應用化學系碩博士班 === 105 === Hepatocellular carcinoma is one of the most serious cancers in Taiwan. People who infect hepatitis B or hepatitis C virus are high risk in developing hepatocellular carcinoma. Since the traditional way of hepatitis B virus examination requires professional technicians and expensive equipment, we aim to develop a feasible and simple method for HBsAg diagnosis. In this study, an ELISA assay is employed for quantitative analysis detection of HBsAg a-determinant. An scFv, obtained from our previous study through a phage display screening, was planed to immobilize on a lab-made NiO chip to function as the capture layer for HBsAg a determinant recognition. However, most of scFvs against a-determinant of HBsAg aggregated to form an inclusion body when they were expressed in E.coli. To improve the solubility of target scFv, the corresponding gene was fused with seven carrier proteins, separately. Among the various tags, Tsf and Bla enhance the expression and solubility of the target scFv. Since the surface of material containing NiO has been shown to possess strong affinity towards protein containing polyhistidine tags, we select NiO foil as the substrate of chip for scFv immobilization and subsequently for HBsAg detection. The NiO foil was prepared through thermal oxidation, which facilitates the formation of metal-oxide nanostructures from metallic materials. By using NiO chip, we successfully demonstrated the sensitivity of HBsAg detection was significantly improved and the detection limit was 34 ng/mL.