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  • 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.

BMC research notes (2017-07-29)
Stephanie P Cartwright, Roslyn M Bill, Bui Tien Sy, Hieu Tran-Van, Hung Minh Nguyen
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Rabbit IgG (whole molecule)–Peroxidase antibody produced in goat, affinity isolated antibody
Sigma-Aldrich
GenElute HP Plasmid Midiprep Kit, sufficient for 25 purifications