Newman Janet, Egan David, Walter Thomas S, Meged Ran, Berry Ian, Ben Jelloul Marouane, Sussman Joel L, Stuart David I, Perrakis Anastassis
Acta Crystallogr D Biol Crystallogr. 2005 Oct;61(Pt 10):1426-31. doi: 10.1107/S0907444905024984. Epub 2005 Sep 28.
A crystallization screening process is presented that was developed for a small academic laboratory. Its underlying concept is to combine sparse-matrix screening with systematic screening in a minimum number of crystallization conditions. The sparse-matrix screen is the cherry-picked combination of conditions from the Joint Center for Structural Genomics (JCSG) extended using conditions from other screens. Its aim is to maximize the coverage of crystallization parameter space with no redundancy. The systematic screen, a pH-, anion- and cation-testing (PACT) screen, aims to decouple the components of each condition and to provide information about the protein, even in the absence of crystals, rather than cover a wide crystallization space. This screening strategy is combined with nanolitre-volume dispensing hardware and a small but practical experiment-tracking system. The screens have been tested both at the NKI and in other laboratories and it is concluded that they provide a useful minimal screening strategy.
本文介绍了一种为小型学术实验室开发的结晶筛选方法。其基本理念是在最少数量的结晶条件下，将稀疏矩阵筛选与系统筛选相结合。稀疏矩阵筛选是从结构基因组学联合中心（JCSG）挑选出的条件组合，并使用其他筛选条件进行了扩展。其目的是在无冗余的情况下最大化结晶参数空间的覆盖范围。系统筛选是一种pH值、阴离子和阳离子测试（PACT）筛选，旨在分离每个条件的组成部分，并即使在没有晶体的情况下也能提供有关蛋白质的信息，而不是覆盖广泛的结晶空间。这种筛选策略与纳升体积分配硬件和一个小型但实用的实验跟踪系统相结合。这些筛选方法已在荷兰癌症研究所（NKI）和其他实验室进行了测试，得出的结论是它们提供了一种有用的最小筛选策略。
