Mariaule Vincent, Dupeux Florine, Márquez José A
European Molecular Biology Laboratory, Grenoble Outstation, Grenoble, France.
Methods Mol Biol. 2014;1091:189-95. doi: 10.1007/978-1-62703-691-7_13.
Construct design and sample formulation are critical in structural biology projects. Large numbers of sample variants are often produced and analyzed for a single target and significant effort is dedicated to sample characterization in order to identify at an early stage the most promising samples to help save manpower and time. Here, we present a method based on a thermal stability assay that can help estimate the likelihood of biological samples to produce crystals. This assay is rapid, inexpensive and consumes very small amounts of sample. The results can be used to prioritize certain constructs at an early stage or as an objective test to help decide when to undertake other type of approaches addressed at improving sample properties.
构建体设计和样品制备在结构生物学项目中至关重要。对于单个靶标,通常会产生并分析大量的样品变体,并且投入大量精力进行样品表征,以便在早期阶段识别出最有前景的样品,从而有助于节省人力和时间。在此,我们提出一种基于热稳定性测定的方法,该方法可帮助估计生物样品产生晶体的可能性。这种测定方法快速、廉价且消耗的样品量非常少。结果可用于在早期阶段对某些构建体进行优先级排序,或作为一种客观测试,以帮助决定何时采用其他类型的方法来改善样品性质。