Xu Xiaowei, Ren Sulin, Chen Xiaoxiao, Ge Jun, Xu Zhenxing, Huang Hongying, Sun Honglin, Gu Yue, Zhou Tong, Li Jianqiang, Xu Hanmei
State Key Laboratory of Nature Medicines, China Pharmaceutical University, Nanjing, 210009, China.
Virol Sin. 2014 Dec;29(6):403-9. doi: 10.1007/s12250-014-3508-9. Epub 2014 Dec 24.
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.
尽管含有乙肝病毒表面抗原（HBsAg）和铝佐剂的传统预防性疫苗早已面市，但仍有近10%的人群无法产生有效的免疫反应。先前的研究表明，乙肝病毒（HBV）PreS2-S在T/B细胞表位中含量丰富，它能诱导比HBsAg更强的免疫反应，尤其是在细胞毒性T淋巴细胞（CTL）反应方面。在本研究中，将编码位于HBsAg N端额外26个氨基酸（PreS2 C端）的HBV PreS2-S基因克隆到pVCH1300表达载体中。在多形汉逊酵母中表达的PreS2-S产量高达250 mg/L。后续的纯化步骤包括对胶体二氧化硅的疏水吸附、离子交换色谱和密度超速离心。最终产物的总收率约为15%，纯度约为99%。与先前的研究一致，使用电子显微镜检测到了约22 nm的病毒样颗粒。所产生的PreS2-S抗原将在动物中进一步研究其有效性和安全性。
