Ge Xin, Filipe Carlos D M
Department of Chemical Engineering, McMaster University, Ontario, Canada.
Biomacromolecules. 2006 Sep;7(9):2475-8. doi: 10.1021/bm060507n.
In this paper, we demonstrate proof-of-principle for a method that allows selective recovery of molecules present at very low concentrations in complex mixtures. The method makes use of an elastin-like polypeptide (ELP) as a coaggregant for the capture of an ELP tagged recombinant protein present at concentrations as low as 10 pM, with a recovery higher than 90%. This coaggregation process was found to be independent of the concentration, at least up to 10 pM concentration of the ELP tagged protein. The coaggregation process is highly specific as was demonstrated by spiking crude cell lysate with the ELP tagged recombinant protein to a final concentration of 1 nM and recovering more than 80% of it to a high level of purity. The method should be particularly useful for high-throughput proteomic studies, where small amounts of poorly expressed proteins could be recovered for analysis by mass spectrometry. In a more general context, the concept presented in this paper provides a method that is highly efficient, specific, and fully reversible, which should render it useful in areas other than recombinant protein purification.
在本文中，我们展示了一种方法的原理验证，该方法能够选择性回收复杂混合物中极低浓度存在的分子。该方法利用一种类弹性蛋白多肽（ELP）作为共聚集剂，用于捕获浓度低至10 pM的ELP标记重组蛋白，回收率高于90%。发现这种共聚集过程与浓度无关，至少在ELP标记蛋白浓度达到10 pM时是这样。共聚集过程具有高度特异性，通过向粗细胞裂解物中加入ELP标记重组蛋白至最终浓度1 nM并将其中80%以上回收至高纯度得以证明。该方法对于高通量蛋白质组学研究应该特别有用，在这类研究中，少量表达不佳的蛋白质可以被回收用于质谱分析。在更一般的背景下，本文提出的概念提供了一种高效、特异且完全可逆的方法，这应该使其在重组蛋白纯化以外的领域也有用途。
