Vero细胞疫苗生产过程中宿主细胞蛋白含量分析

Analysis of Host Cell Protein Content During Production of Vero Cell-derived Vaccine

目的了解Vero细胞疫苗中间产品中宿主细胞蛋白(Host cell protein,HCP)的含量变化。方法模拟Vero细胞疫苗生产工艺流程,制备Vero细胞HCP,免疫家兔制备Vero细胞HCP免疫血清,经Protein ASepharose CL-4B亲和层析,制备纯化的抗Vero细胞HCPIgG,通过SDS-PAGE和Western blot分析Vero细胞乙型脑炎疫苗及Vero细胞SARS疫苗各中间产品的蛋白、HCP及病毒抗原成分的含量变化。结果Vero细胞乙型脑炎疫苗和Vero细胞SARS疫苗中间产品中均存在一定量HCP,不同的纯化工艺均可以去除大部分HCP,随着生产工艺的进行,疫苗中间产品中HCP的含量逐渐降低,病毒成分的纯度逐渐增高,但纯化后产品中仍存在少量HCP。结论Vero细胞乙型脑炎疫苗和Vero细胞SARS疫苗均存在一定量HCP。

Objective To explore the change of host cell protein（HCP） content in the intermediate product during production of Veto cell-derived vaccine. Methods Prepare the HCP of Vero cells by imitating the production process of Vero cell-derived vaccine. Immunize rabbit with the HCP to obtain immune sera, and prepare purified IgG against Vero cell HCP by Protein A Sepharose CL-4B affinity chromatography. Analyze the changes of protein, HCP and virus antigen contents in intermediate product during production of Vero cell-derived Japanese encephalifis（JE） vaccine and Vero cell-derived SAILS vaccine by SDS-PAGE and Western blot.Results HCPs were observed in both the intermediate product during production of Vero cell-derived Japanese encephalitis vaccine and Vero cell-derived SARS vaccine. Most HCP might be removed by purification process. During production, the HCP in intermediate product decreased gradually, and the purity of virus component increased gradually. However, a small quantity of HCP still existed in the puff.fled product. Conclusion The HCP existed in both JE and SARS vaccines prepared with Vero cells.