Lu Xue Xin, Yi Yao, Su Qiu Dong, Bi Sheng Li
National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Biomed Environ Sci. 2016 Jun;29(6):417-23. doi: 10.3967/bes2016.054.
Hepatitis Delt a Virus (HDV) antigen is widely used as a capture antigen in ELISAs for the identification of HDV infection; large amounts of recombinant HDV antigen with active antigenicity are required for this purpose.
Reconstruct the gene of HDV antigen based on the bias code of Escherichia coli, the recombinant protein expresses by high-density fermentation with fed-batch feeding strategy, and purify by immobilized metal chromatography. The sensitivity and specificity of this antigen detect by ELISA method.
The expression of HDV antigen can reach 20% of the total cell mass in the soluble form. The recombinant HDV antigen can be conveniently purified (98%) by immobilized metal ion affinity chromatography (IMAC) using the interaction between a His-tag and nickel ions. Production of recombinant HDV antigen can reach 0.5 g/L under conditions of high-density cell fermentation. Applied to the diagnostic ELISA method, the recombinant HDV antigen shows excellent sensitivity (97% for IgM and 100% for IgG) and specificity (100% for IgG and IgM) for the detection of anti-HDV antibodies.
Expression and purification the recombinant HDV antigen as a candidate protein for application in a diagnostic ELISA for HDV infection. Large-scale production of the protein can be achieved using the high-density fermentation strategy.
丁型肝炎病毒（HDV）抗原在用于鉴定HDV感染的酶联免疫吸附测定（ELISA）中被广泛用作捕获抗原；为此需要大量具有活性抗原性的重组HDV抗原。
基于大肠杆菌的偏好密码子重建HDV抗原基因，采用补料分批培养策略通过高密度发酵表达重组蛋白，并通过固定化金属色谱法进行纯化。采用ELISA法检测该抗原的敏感性和特异性。
HDV抗原以可溶形式表达量可达细胞总质量的20%。利用His标签与镍离子之间的相互作用，通过固定化金属离子亲和色谱法（IMAC）可方便地纯化重组HDV抗原（纯度达98%）。在高密度细胞发酵条件下，重组HDV抗原产量可达0.5 g/L。应用于诊断ELISA法时，重组HDV抗原在检测抗HDV抗体方面显示出优异的敏感性（IgM为97%，IgG为100%）和特异性（IgG和IgM均为100%）。
表达并纯化重组HDV抗原，作为用于HDV感染诊断ELISA的候选蛋白。采用高密度发酵策略可实现该蛋白的大规模生产。
