摘要
The baculovirus system has the potential for the large-scale production of protein products by two methods. First, as a result of recent advances (1,5), large-scale cell culture is now possible and, second, the recombinant baculovirus can be used to infect susceptible insect larvae (3). Whether using small-scale cell culture or the aforementioned methods for scale-up, the requirement for downstream processing of the protein product is manifest. Purification of the product is greatly facilitated when expression is high (ca. 30% of total cell protein); however, when expression is relatively low or membrane proteins are required, the researcher faces the same problems encountered in ascertaining the optimum conditions for any de novo protein purification. Obviously, the degree of purity required for a given product is indicative of its final use; consequently, a number of antigens for use in diagnostic procedures have been relatively crude preparations (4,5). This chapter highlights the methods that have been used to purify baculovirusexpressed protein products, and is aimed at providing general guidelines to the purification of certain types of protein product rather than a definitive guide to protein purification.
摘要译文
杆状病毒系统具有通过两种方法大规模生产蛋白质产品的潜力。首先，作为最近的进展（1,5）的结果，现在可以进行大规模细胞培养，其次，重组杆状病毒可以用于感染易感昆虫幼虫（3）。无论是使用小规模细胞培养还是上述扩大规模的方法，都显示出对蛋白质产物的下游加工的要求。当表达高时，大大促进了产物的纯化（大约总细胞蛋白质的30％）;然而，当表达相对较低或需要膜蛋白时，研究人员面临着在确定任何从头蛋白纯化的最佳条件时遇到的相同问题。显然，给定产品所需的纯度表明其最终用途;因此，用于诊断程序的许多抗原是相对粗制的制剂（4,5）。本章重点介绍了用于纯化杆状病毒表达的蛋白质产品的方法，旨在为某些类型的蛋白质产品的纯化提供一般指导，而不是蛋白质纯化的权威指南。
Vincent C. Emery[1]. Baculovirus Expression Vectors. Practical Molecular Virology[M].DE: Springer, 1992: 319-326