Separating full-length protein from aggregating proteolytic products using filter flow-through purification
Separating full-length protein from aggregating proteolytic products using filter flow-through purification
复制标题
使用过滤流通纯化从聚集的蛋白水解产物中分离全长蛋白质
DOI：
10.1016/j.ab.2016.09.009
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发表时间：
2016
影响因子：
2.9
通讯作者：
Bondos, Sarah E.
中科院分区：
文献类型：
--
作者：
Churion, Kelly A.;Rogers, Robert E.;Bayless, Kayla J.;Bondos, Sarah E.
从蛋白水解物中分离全长蛋白质是一项具有挑战性的工作，因为分离蛋白质所用的特性也可能存在于蛋白水解物中。许多分离技术存在非特异性蛋白质黏附的风险，和/或需要大量时间，使蛋白质在裂解后能够继续分解和聚集。我们证明了两种不同蛋白质的蛋白质分解产物聚集在一起。因此，通过过滤流过纯化，可以从这些片段中快速分离出全长蛋白质，从而大幅提高蛋白质纯度。这种快速的方法可能对内在无序的蛋白质有用，因为其重复的序列组成和灵活的性质可以促进聚集。
Separation of full-length protein from proteolytic products is challenging, since the properties used to isolate the protein can also be present in proteolytic products. Many separation techniques risk non-specific protein adhesion and/or require a lot of time, enabling continued proteolysis and aggregation after lysis. We demonstrate that proteolytic products aggregate for two different proteins. As a result, full-length protein can be rapidly separated from these fragments by filter flow-through purification, resulting in a substantial protein purity enhancement. This rapid approach is likely to be useful for intrinsically disordered proteins, whose repetitive sequence composition and flexible nature can facilitate aggregation.