Zhang Xiao, Wei Minxi, Pan Huirong, Lin Zhijie, Wang Kaihang, Weng Zusen, Zhu Yibin, Xin Lu, Zhang Jun, Li Shaowei, Xia Ningshao, Zhao Qinjian
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiamen, Fujian 361005, PR China; School of Public Health, Xiamen University, Xiamen, Fujian 361005, PR China.
State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, National Institute of Diagnostics and Vaccine Development in Infectious Diseases, Xiamen University, Xiamen, Fujian 361005, PR China; School of Life Science, Xiamen University, Xiamen, Fujian 361005, PR China.
Vaccine. 2014 Jul 7;32(32):4039-50. doi: 10.1016/j.vaccine.2014.05.064. Epub 2014 Jun 2.
The hepatitis E virus (HEV) vaccine, Hecolin(®), was licensed in China for the prevention of HEV infection and HEV-related diseases with demonstrated safety and efficacy [1,2]. The vaccine is composed of a truncated HEV capsid protein, p239, as the sole antigen encoded by open reading frame 2 and produced using Escherichia coli platform. The production of this virus-like particle (VLP) form of the antigen was successfully scaled up 50-fold from a bench scale to a manufacturing scale. Product consistency was demonstrated using a combination of biophysical, biochemical and immunochemical methods, which revealed comparable antigen characteristics among different batches. Particle size of the nanometer scale particulate antigen and presence of key epitopes on the particle surface are two prerequisites for an efficacious VLP-based vaccine. The particle size was monitored by several different methods, which showed diameters between 20 and 30nm for the p239 particles. The thermal stability and aggregation propensity of the antigen were assessed using differential scanning calorimetry and cloud point assay under heat stress conditions. Key epitopes on the particulate antigen were analyzed using a panel of murine anti-HEV monoclonal antibodies (mAbs). The immuno reactivity to the mAbs among the different antigen lots was highly consistent when analyzed quantitatively using a surface plasmon resonance technique. Using a sandwich ELISA to probe the integrity of two different epitopes in the antigen, the specific antigenicity of multiple batches was assessed to demonstrate consistency in these critical product attributes. Overall, our findings showed that the antigen production process is robust and scalable during the manufacturing of Hecolin(®).
戊型肝炎病毒（HEV）疫苗“益可宁”（Hecolin®）已在中国获批用于预防HEV感染及HEV相关疾病，且已证实其安全性和有效性[1,2]。该疫苗由截短的HEV衣壳蛋白p239组成，它是开放阅读框2编码的唯一抗原，采用大肠杆菌表达平台生产。这种抗原的病毒样颗粒（VLP）形式的产量已成功从实验室规模扩大了50倍，达到生产规模。通过生物物理、生化和免疫化学方法相结合证明了产品的一致性，结果显示不同批次之间的抗原特性具有可比性。纳米级颗粒抗原的粒径以及颗粒表面关键表位的存在是基于VLP的有效疫苗的两个先决条件。通过几种不同方法监测粒径，结果显示p239颗粒的直径在20至30nm之间。在热应激条件下，使用差示扫描量热法和浊点测定法评估抗原的热稳定性和聚集倾向。使用一组鼠抗HEV单克隆抗体（mAb）分析颗粒抗原上的关键表位。当使用表面等离子体共振技术进行定量分析时，不同抗原批次对mAb的免疫反应性高度一致。使用夹心ELISA检测抗原中两个不同表位的完整性，评估多个批次的特异性抗原性，以证明这些关键产品属性的一致性。总体而言，我们的研究结果表明，在“益可宁”（Hecolin®）的生产过程中，抗原生产工艺稳健且可扩展。
