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Protein Crystallization Screening Using Associative Experimental Design

机译:使用相关实验设计的蛋白质结晶筛选

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Protein crystallization remains a highly empirical process. The purpose of protein crystallization screening is the determination of the main factors of importance leading to protein crystallization. One of the major problems about determining these factors is that screening is often expanded to many hundreds or thousands of conditions to maximize combinatorial chemical space coverage for a successful (crystalline) outcome. In this paper, we propose a new experimental design method called "Associative Experimental Design (AED)" that provides a list of screening factors that are likely to lead to higher scoring outcomes or crystals by analyzing preliminary experimental results. We have tested AED on Nucleoside diphosphate kinase, HAD superfamily hydrolase, and nucleoside kinase proteins derived from the hyperthermophile Ther-mococcus thioreducens. After obtaining the candidate novel conditions, we have confirmed that AED method yielded high scoring crystals after experimenting in a wet lab.
机译:蛋白质结晶仍然是一个高度经验的过程。蛋白质结晶筛选的目的是确定导致蛋白质结晶的重要因素。确定这些因素的主要问题之一是,筛选通常会扩展到数百或数千个条件,以最大化组合化学空间的覆盖范围,以获得成功的(晶体)结果。在本文中,我们提出了一种称为“关联实验设计(AED)”的新实验设计方法,该方法通过分析初步实验结果,提供了可能导致更高得分结果或晶体的筛选因素列表。我们已经对核苷二磷酸激酶,HAD超家族水解酶和衍生自嗜热嗜热球菌硫还原菌的核苷激酶蛋白进行了AED测试。在获得候选新条件后,我们已经确认,在湿实验室中进行实验后,AED方法可产生高分晶体。

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