title: Real-time stability of a hepatitis E vaccine (Hecolin®) demonstrated with potency assays and multifaceted physicochemical methods.
abstract: The first prophylactic vaccine against hepatitis E virus (HEV), Hecolin®, was licensed in China. Recombinant p239 virus-like particle (VLP) is its active component with dimeric protein as the basic building block harboring the immuno dominant and neutralizing epitopes. The real time and real condition stability of the prefilled syringes for the vaccine was demonstrated using both in vivo mouse potency and in vitro antigenicity assays. A total of 12 lots of Hecolin® were assessed with a set of assays after storage at 2-8°C for 24months. The particle characteristics of p239 VLP recovered from the aluminum-containing adjuvant was assessed with different methods including analytical ultracentrifugation, high performance size exclusion chromatography and transmission electron microscopy. The thermal and conformational stability of the adsorbed antigen was assessed using differential scanning calorimetry. The protein integrity of the recovered p239 antigen was demonstrated using SDS-PAGE with silvering staining, LC-MS and MALDI-TOF MS. Most importantly, the binding activity to the neutralizing antibody or vaccine antigenicity was measured using an epitope-specific and real-time SPR assay and a monoclonal antibody-based sandwich ELISA. Taken together, the overall good stability of the Hecolin® prefilled syringes was demonstrated with unaltered molecular and functional attributes after storage at 2-8°C for 24months.
authors: 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.
authors: Xiao
authors: X
authors: Zhang
authors: Xiamen Innovax Biotech Co., Ltd, Xiamen, Fujian 361005, PR China.
authors: Minxi
authors: M
authors: Wei
authors: Xiamen Innovax Biotech Co., Ltd, Xiamen, Fujian 361005, PR China.
authors: Guang
authors: G
authors: Sun
authors: 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.
authors: Xin
authors: X
authors: Wang
authors: 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.
authors: Min
authors: M
authors: Li
authors: Xiamen Innovax Biotech Co., Ltd, Xiamen, Fujian 361005, PR China.
authors: Zhijie
authors: Z
authors: Lin
authors: Xiamen Innovax Biotech Co., Ltd, Xiamen, Fujian 361005, PR China.
authors: Zhongyi
authors: Z
authors: Li
authors: 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.
authors: Yufang
authors: Y
authors: Li
authors: 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.
authors: Mujin
authors: M
authors: Fang
authors: 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.
authors: Jun
authors: J
authors: Zhang
authors: 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; School of Life Science, Xiamen University, Xiamen, Fujian 361005, PR China.
authors: Shaowei
authors: S
authors: Li
authors: 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; School of Life Science, Xiamen University, Xiamen, Fujian 361005, PR China. Electronic address: nsxia@xmu.edu.cn.
authors: Ningshao
authors: N
authors: Xia
authors: 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. Electronic address: qinjian_zhao@xmu.edu.cn.
authors: Qinjian
authors: Q
authors: Zhao
mesh_terms: D000276:Adjuvants, Immunologic; D000818:Animals; D057134:Antibodies, Neutralizing / Q000276:immunology; D002152:Calorimetry, Differential Scanning; D002681:China / Q000453:epidemiology; D003080:Cold Temperature; D004356:Drug Storage; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D006508:Hepatitis Antibodies / Q000276:immunology; D016751:Hepatitis E / Q000453:epidemiology / Q000517:prevention & control; D016752:Hepatitis E virus / Q000276:immunology; D000071497:Immunogenicity, Vaccine; D051379:Mice; D064166:Vaccine Potency; D014614:Vaccines, Synthetic / Q000737:chemistry* / Q000276:immunology*; D058425:Vaccines, Virus-Like Particle / Q000737:chemistry / Q000276:immunology; D014761:Viral Hepatitis Vaccines / Q000737:chemistry* / Q000276:immunology*
chemical_list: D000276:Adjuvants, Immunologic; D057134:Antibodies, Neutralizing; D000939:Epitopes; D006508:Hepatitis Antibodies; D014614:Vaccines, Synthetic; D058425:Vaccines, Virus-Like Particle; D014761:Viral Hepatitis Vaccines; C577000:hecolin
keywords: Hepatitis E vaccine; Hepatitis E virus; In vitro relative potency (IVRP); In vivo mouse potency; Monoclonal antibody; Neutralizing epitope; Real-time stability