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

Stephanie Cartwright, Roslyn M. Bill, Sy Tien Bui, Hieu Tran Van, Hung Minh Nguyen*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Article number340
Number of pages6
JournalBMC Research Notes
Volume10
DOIs
Publication statusPublished - 27 Jul 2017

Fingerprint

Hepatitis delta Antigens
Hepatitis Delta Virus
Pichia
Viruses
Yeast
Purification
Yeasts
Antigens
Virus Diseases
Vaccines
Gene encoding
Hepatitis B
Liver Cirrhosis
Liver
Hepatitis
Methanol
Hepatocellular Carcinoma
Assays
Protein Isoforms
Infection

Bibliographical note

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Keywords

  • HDAg
  • hepatitis delta virus
  • pichia pastoris
  • protein expression

Cite this

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abstract = "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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Rapid expression and purification of the hepatitis delta virus antigen using the methylotropic yeast Pichia pastoris. / Cartwright, Stephanie; Bill, Roslyn M.; Bui, Sy Tien; Van, Hieu Tran; Nguyen, Hung Minh.

In: BMC Research Notes, Vol. 10, 340, 27.07.2017.

Research output: Contribution to journalArticle

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