Protein Purification
Protein Purification
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DOI：
10.2174/157341109787846171
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发表时间：
2009-04-01
影响因子：
1.8
通讯作者：
Swiatek, Gavin
中科院分区：
文献类型：
--
作者：
Ward, William W.;Swiatek, Gavin
蛋白质纯化的科学、艺术和实践已经伴随我们超过了一个世纪，然而，在许多方面，该领域只是现在才发展过去的青少年根源。新方法正在以令人眼花缭乱的速度取代旧方法，即使是终身蛋白质纯化专家也无法跟上。在这篇文章中，我们提出了许多国家的最先进的蛋白质纯化技术，而不完全忽视过去。我们的目标是使那些相对较新的蛋白质纯化领域选择最好的方法来解决自己的纯化问题。我们所描述的每一种方法都在我们自己的研究中得到了使用和验证。我们描述这些方法，指出优点，缺点和局限性与实际的例子，而不是复杂的，理论方程。本文涵盖了提取，澄清，批量纯化，低压柱层析，HPLC和电泳的方法，适用于基因工程，重组蛋白质和蛋白质分离的天然来源。比较新的方法，三相分配，疏水电荷诱导色谱，固定化金属亲和色谱，和灌注色谱的特点。
The science, art, and practice of protein purification have been with us for more than a century, yet, in many respects, the field is only now evolving past its adolescent roots. New methods are replacing old methods at such a dizzying pace, that even life-long experts in protein purification cannot keep up. In this article, we present many state-of-the-art protein purification techniques without totally ignoring the past. Our goal is to enable those relatively new to the field of protein purification to choose the best methods to solve their own purification problems. Each method we describe has been used and validated in our own research. We describe these methods, pointing out advantages, disadvantages, and limitations with practical examples rather than with complex, theoretical equations. This paper covers methods of extraction, clarification, batch purification, low pressure column chromatography, HPLC, and electrophoresis as applied to both genetically engineered, recombinant proteins and proteins isolated from natural sources. The relatively new methods of three-phase partitioning, hydrophobic charge induction chromatography, immobilized metal affinity chromatography, and perfusion chromatography are featured.