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篇名 同步輻射蛋白質結晶學之高通量暨高純度蛋白質純化設施應用
卷期 30:5=169
並列篇名 Automation of High-Throughput Protein Purification for X-Ray Crystallography Study
作者 劉思恒黃詩芳王聖宏潘冠宇林易弘
頁次 24-31
出刊日期 200904

中文摘要

同步輻射在蛋白質結構生物學上的應用與研究發展至今,已是一項非常成熟的技術。以X光繞射為工具,在原子解析度下,正確地鑑定並提供詳實的蛋白質晶體結構,以探討蛋白分子結構與功能的關係,有助於基礎生命科學的發展。研究結構基因體學過程中最大的挑戰在於大量(high-throughput)及同時並行(parallel)多種基因所相對應的蛋白質表現、純化及其結構的確定與分析。在蛋白質純化方面,主要的考量在於如何快速獲得大量高純度且溶解度高的蛋白質,以因應篩選高品質蛋白晶體時的大量需求與純度需要。因此,建立AKTAexplorer3D以及AKTAxpress之高效能蛋白質自動純化裝置工作站,將會是解決此一瓶頸的最佳利器。AKTAxpress即專為純化大量且高純度重組蛋白所重新設計研發的自動化純化系統。

英文摘要

Advances in genomics research provide valuable information about the composition of proteins, but little about their structures and most crucially, little concerning their functions. Thus, it is essential to know their three-dimensional structures and further to understand how and why proteins function as they do. To date, structural genomics is a field of research that has been established in particular for deciphering of the 3-D structure of proteins via a high-throughput methodologies and techniques. The critical challenge in the studies of structural genomics in general, is the availability of milligram-quantity and most importantly high-quality soluble monodisperse proteins. The choice of protein purification and handling procedures plays a critical role in obtaining high-quality proteins. In this regard, we introduce AKTA explorer 3D and AKTAxpress workstations that has been successfully applied to many structural genomics centers, capable of performing multidimensional chromatography. Various purification strategies, their design and their implementation are discussed in this study.

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