细胞制备流行性感冒病毒疫苗

Advance in Cell derived influenza virus vaccine process development

关键词:  细胞流感疫苗  研究进展

Key words:  cell-derived influenza vaccine  process development

摘要:

目前，流行性感冒病毒疫苗（流感疫苗）的生产大多采用鸡胚，这种传统的方法劳动强度大，鸡胚用量大，需要繁琐的纯化工艺去除鸡胚蛋白减少过敏反应，流感的大流行性以及新发流感的流行迫切需要一种新的疫苗生产方法：用哺乳动物细胞生产疫苗。细胞生产流感疫苗经过研究和发展，从可用于生产的Vero、MDCK、PEG-C6细胞的生物学鉴定、分析到流感病毒的重配、适用、培养以及下游工艺的悬浮培养、浓缩、纯化、安全以及效果观察都取得长足进展。本文从细胞流感疫苗的细胞基质、病毒适用、重配、培养、浓缩、纯化及其疫苗的安全、效果观察方面综述了细胞流感疫苗的研发重点。

Abstract:

This manuscript discusses the mammalian cell line used to grow influenza virus adapted cultivation, as well as downstream process development, suspension cultivation, concentration, purification for influenza vaccines. Influenza viruses for production are presently produced in embryonated hen’s eggs. This conventional standard methodology is extremely cumbersome; it requires millions of eggs and an extensive purification to reduce the amount of contaminating egg proteins and to minimise the risk of allergies against egg albumin. The shortage of eggs in a pandemic situation, the selection of egg-adapted variants and the presence of adventitious viruses has emphasised the necessity for production of Influenza vaccines on a well characterised stable mammalian cell line. Mammalian cell-derived vaccines, produced in nontumorigenic cell lines, have been developed and shown to be effective production systems as alternatives to eggs. Established Vero, MDCK, PER-C6 technology has been successfully adapted to isolation , reassortant virus with reverse genetics to generate high growth virus strain as well as to large scale production of a huge variety of Influenza virus strains. The production in 1~1200 liter fermenter cultures under serum free conditions gave antigen yields comparable to the conventional embryonated egg technology. The development of a rapid and efficient purification scheme resulted in a safe high purity vaccine which was at least as immunogenic as conventional egg-derived vaccines in a mouse model. Clinical trials in different region demonstrated that the cell-derived influenza vaccine is well tolerated, safe and highly immunogenic in humans.