Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Postal 510-3, Cuernavaca, Morelos, CP 62250, Mexico.
Methods. 2012 Mar;56(3):389-95. doi: 10.1016/j.ymeth.2012.01.004. Epub 2012 Jan 28.
The insect cell-baculovirus system (IC-BEVS) is widely used for the production of recombinant viral proteins for vaccine applications. It is especially suitable for the production of virus-like particles, which often require the simultaneous production of several recombinant proteins. Here, the available tools and process requirements for the simultaneous production of several recombinant proteins using the IC-BEVS are discussed. The production of double-layered rotavirus like particles is used as a specific example for the simultaneous production of two recombinant proteins. Methods to quantify VLP in small samples are described. The multiplicity and time of infection are presented as tools to manipulate protein concentration, and the effect on protein concentration ratios on the assembly efficiency of double-layered rotavirus like particles is discussed. It was found that not only the ratio between the recombinant proteins is determinant of VLP assembly efficiency, but also that assembly efficiency is related to the characteristics of the assembled proteins. This is the first time that kinetics of VLP production are followed during cultures, and that the assembly efficiency is quantitatively determined.
昆虫细胞-杆状病毒系统（IC-BEVS）广泛用于生产用于疫苗应用的重组病毒蛋白。它特别适合生产病毒样颗粒（VLPs），通常需要同时生产几种重组蛋白。在这里，讨论了使用 IC-BEVS 同时生产几种重组蛋白的可用工具和工艺要求。双层轮状病毒样颗粒的生产用作同时生产两种重组蛋白的具体示例。还描述了用于在小样本中定量 VLP 的方法。感染的多重性和时间被用作操纵蛋白浓度的工具，并讨论了对双层轮状病毒样颗粒组装效率的蛋白浓度比的影响。结果发现，不仅重组蛋白之间的比率决定了 VLP 组装效率，而且组装效率与组装蛋白的特性有关。这是首次在培养过程中跟踪 VLP 生产的动力学，并定量确定组装效率。
