Rapid expression and purification of the hepatitis delta virus antigen using the methylotropic yeast Pichia pastoris.
Rapid expression and purification of the hepatitis delta virus antigen using the methylotropic yeast Pichia pastoris.
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使用甲基酵母pichia pardoris的肝炎三角病病毒抗原的快速表达和纯化。
DOI：
10.1186/s13104-017-2692-8
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发表时间：
2017-07-27
影响因子：
1.8
通讯作者：
Nguyen HM
中科院分区：
文献类型：
--
作者：
Cartwright SP;Bill RM;Sy BT;Tran-Van H;Nguyen HM
与单一病毒感染的患者相比，双重B型肝炎（HBV）和丁型肝炎（HDV）病毒感染的患者进展为肝硬化和肝细胞癌的风险增加。治疗由HBV/HDV双重感染引起的病毒性肝炎是一个挑战。目前还没有针对HDV的疫苗。HDV抗原（HDAg）的重组生产是迈向潜在疫苗候选物和HDV检测方法开发的第一步。本研究证明了HDAg同种型S-HDAg在毕赤酵母中的表达。设计了一个重组载体，该载体携带在甲醇诱导型启动子AOX 1控制下的编码S-HDAg的标记基因，并整合到巴斯德毕赤酵母X33中。该蛋白，这是纯化使用Ni 2+亲和柱和洗脱在100-150 mM咪唑，有潜力作为重组抗原进行进一步研究。
Patients with dual hepatitis B (HBV) and hepatitis D (HDV) virus infection are at an increased risk of progression to liver cirrhosis and hepatocellular carcinoma than patients with a single viral infection. Treatment of viral hepatitis due to dual HBV/HDV infection represents a challenge. Currently there is no vaccine against HDV. Recombinant production of HDV antigen (HDAg) is the first step towards a potential vaccine candidate and the development of assays for HDV detection. This study demonstrates the expression of one HDAg isoform, S-HDAg, in Pichia pastoris. A recombinant vector carrying a tagged gene encoding S-HDAg under the control of the methanol-inducible promoter AOX1 was designed and integrated into P. pastoris X33. The protein, which was purified using a Ni2+ affinity column and eluted at 100–150 mM imidazole, has potential as a recombinant antigen for further study.