大规模培养及浓缩爱泼斯坦-巴尔病毒的新方法。
New method for large-scale growth; and concentration of the Epstein-Barr viruses.
作者信息
Shibley G P, Manousos M, Munch K, Zelljadt I, Fisher L, Mayyasi S, Harewood K, Stevens R, Jensen K E
出版信息
Appl Environ Microbiol. 1980 Dec;40(6):1044-8. doi: 10.1128/aem.40.6.1044-1048.1980.
Efficacious systems are described for the large-scale growth in tissue culture and concentration of infectious (P3HR-1) and transforming (B95-8) Epstein-Barr virus. Also recorded here are our updated procedures for growing stock cultures and protocols to harvest fluids containing biologically active virus which is infectious or transforming. Various methods of concentrating biologically active Epstein-Barr virus have been evaluated. Cellular debris can be removed efficiently and rapidly from culture harvest fluids by clarification through a JCF-Z continuous-flow rotor. Efficient and reliable virus concentration was achieved by molecular filtration with Millipore Pellicon cassettes, using flow rates to 10 liters/h to produce fivefold concentrates followed by pelletization in a fixed-angle rotor. Data from recent production lots showed an average infectivity titer for P3HR-1 virus of 10(4.5) early antigen units per ml (100-fold concentrate) and 10(5.7) transforming units per ml (200-fold concentrate) for B95-9 virus lots.
本文描述了用于在组织培养中大规模培养以及浓缩感染性（P3HR-1）和转化性（B95-8）爱泼斯坦-巴尔病毒的有效系统。这里还记录了我们用于培养储备培养物的更新程序以及收获含有具有感染性或转化性的生物活性病毒的液体的方案。已经评估了多种浓缩生物活性爱泼斯坦-巴尔病毒的方法。通过使用JCF-Z连续流转子进行澄清，可以高效、快速地从培养收获液中去除细胞碎片。通过使用密理博Pellicon盒式滤器进行分子过滤实现了高效且可靠的病毒浓缩，流速可达10升/小时，以产生五倍浓缩物，随后在固定角度转子中进行沉淀。近期生产批次的数据显示，P3HR-1病毒的平均感染滴度为每毫升10(4.5)个早期抗原单位（100倍浓缩物），B95-9病毒批次每毫升为10(5.7)个转化单位（200倍浓缩物）。
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