Rapid expression and purification of the hepatitis delta virus antigen using the methylotropic yeast Pichia pastoris

利用甲基营养酵母毕赤酵母快速表达和纯化丁型肝炎病毒抗原

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作者:Stephanie P Cartwright, Roslyn M Bill, Bui Tien Sy, Hieu Tran-Van, Hung Minh Nguyen

Objective

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.

Results

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.

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