title: Asymmetrical flow field-flow fractionation coupled with multi-angle laser light scattering for stability comparison of virus-like particles in different solution environments.
abstract: The stabilities of two commercially available virus like particles, CHO-HBsAg expressed by Chinese hamster ovary (CHO) cells and Hans-HBsAg expressed by Hansenula polymorpha (Hans), were compared using AF4-MALLS under different treatment processes. The initial molecular weight and hydrodynamic diameter of CHO-HBsAg measured with AF4-MALLS were 4727kDa and 29.4nm, while those of Hans-HBsAg were 3039kDa and 22.8nm respectively. In salt solution of 2M ammonium sulfate, the molecular weight and size of CHO-HBsAg had little change, and its antigenicity remained 95%, while those of Hans-HBsAg changed greatly, resulting in aggregation and 75% antigenicity loss. In freeze-thaw operations, Hans-HBsAg aggregated heavily. Most of the aggregates precipitated and the rest soluble aggregates reached 10(5)-10(6)kDa in molecular weight. The antigenicity of Hans-HBsAg decreased to 26.9% after five freeze-thaw cycles. For CHO-HBsAg, there was no obvious aggregation in freeze-thaw, and the antigenicity retained above 98%. In heating process, Hans-HBsAg gradually aggregated to large particles with temperature and the antigenicity decreased to 10% when the temperature reached 80°C. In contrast, CHO-HBsAg would not aggregate with temperature, remained 92% antigenicity at 80°C. The study demonstrated that CHO-HBsAg appeared to be a superior vaccine antigen in term of particle stability and constant antigenicity, which are important in production, transportation and storage.
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Chen
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China.
authors.forename: Yuefang
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China.
authors.forename: Jian
authors.initials: J
authors.lastname: Luo
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China; Jiangsu National Synergetic Innovation Center for Advanced Materials (SICAM), Nanjing 210023, PR China. Electronic address: zgsu@ipe.ac.cn.
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Su
mesh_terms: D000818:Animals; D016466:CHO Cells; D003412:Cricetulus; D031521:Fractionation, Field Flow; D005615:Freezing; D006514:Hepatitis B Surface Antigens / Q000737:chemistry*; D008970:Molecular Weight; D010316:Particle Size; D010843:Pichia; D066329:Protein Aggregates; D055550:Protein Stability; D013696:Temperature; D058425:Vaccines, Virus-Like Particle / Q000737:chemistry*
chemical_list: D006514:Hepatitis B Surface Antigens; D066329:Protein Aggregates; D058425:Vaccines, Virus-Like Particle
keywords: Aggregation; Antigenicity; Asymmetrical flow field-flow fractionation; Hepatitis B surface antigen; Virus-like particle