title: Generation of hepatitis B virus PreS2-S antigen in Hansenula polymorpha.
abstract: Despite the long availability of a traditional prophylactic vaccine containing the HBV surface antigen (HBsAg) and aluminum adjuvant, nearly 10% of the population remains unable to generate an effective immune response. Previous studies have indicated that hepatitis B virus (HBV) PreS2-S is abundant in T/B cell epitopes, which induces a stronger immune response than HBsAg, particularly in terms of cytotoxic T lymphocyte (CTL) reaction. In the current study, the HBV PreS2-S gene encoding an extra 26 amino acids (PreS2 C-terminus) located at the N-terminus of HBsAg was cloned into the pVCH1300 expression vector. PreS2-S expressed in the methylotrophic yeast, Hansenula polymorpha, was produced at a yield of up to 250 mg/L. Subsequent purification steps involved hydrophobic adsorption to colloidal silica, ion-exchange chromatography and density ultracentrifugation. The final product was obtained with a total yield of ∼ 15% and purity of ∼ 99%. In keeping with previous studies, ∼ 22 nm viruslike particles were detected using electron microscopy. The generated PreS2-S antigen will be further studied for efficacy and safty in animals.
authors.affiliation: State Key Laboratory of Nature Medicines, China Pharmaceutical University, Nanjing, 210009, China.
authors.forename: Xiaowei
authors.initials: X
authors.lastname: Xu
authors.forename: Sulin
authors.initials: S
authors.lastname: Ren
authors.forename: Xiaoxiao
authors.initials: X
authors.lastname: Chen
authors.forename: Jun
authors.initials: J
authors.lastname: Ge
authors.forename: Zhenxing
authors.initials: Z
authors.lastname: Xu
authors.forename: Hongying
authors.initials: H
authors.lastname: Huang
authors.forename: Honglin
authors.initials: H
authors.lastname: Sun
authors.forename: Yue
authors.initials: Y
authors.lastname: Gu
authors.forename: Tong
authors.initials: T
authors.lastname: Zhou
authors.forename: Jianqiang
authors.initials: J
authors.lastname: Li
authors.forename: Hanmei
authors.initials: H
authors.lastname: Xu
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D015870:Gene Expression; D006514:Hepatitis B Surface Antigens / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics* / Q000378:metabolism; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011498:Protein Precursors / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D006514:Hepatitis B Surface Antigens; D011498:Protein Precursors; C050807:presurface protein 2, hepatitis B surface antigen