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首页> 外文期刊>Journal of chromatography, A: Including electrophoresis and other separation methods >Nickel(II)-immobilized sulfhydryl cotton fiber for selective binding and rapid separation of histidine-tagged proteins
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Nickel(II)-immobilized sulfhydryl cotton fiber for selective binding and rapid separation of histidine-tagged proteins

机译:镍(II)固定的巯基棉纤维用于组氨酸标签蛋白的选择性结合和快速分离

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摘要

In the current study, a novel nickel(II)-immobilized sulfhydryl cotton fiber (SCF-Ni2+) was prepared in a simple way based on the coordination effect between Ni2+ and thiol group on the surface of SCF. The composition and element mapping of SCF-Ni2+ fibers were demonstrated by energy-dispersive X-ray (EDX) spectroscopy. Based on the high affinity of Ni2+ to 6 x His on histidine-tagged (His-tagged) proteins, SCF-Ni2+ fibers were then further used as an immobilized metal ion affinity chromatography (IMAC) adsorbent for selective binding and rapid separation of His-tagged proteins using an in- pipette-tip SPE format. Our results showed that SCF-Ni2+ adsorbent can selectively capture His-tagged proteins from protein mixture and Escherichia coli cell lysates. Taken together, the developed method provides a rapid, convenient and efficient approach for the purification of His-tagged proteins. (C) 2015 Elsevier B.V. All rights reserved.
机译:在当前的研究中,基于Ni 2+和SCF表面硫醇基团之间的配位作用,以简单的方式制备了新型的镍(II)固定的巯基棉纤维(SCF-Ni 2+)。 SCF-Ni2 +纤维的组成和元素图谱通过能量色散X射线(EDX)光谱证实。基于Ni2 +对组氨酸标签的(His标签)蛋白具有6 x His的高亲和力,SCF-Ni2 +纤维随后被进一步用作固定的金属离子亲和色谱(IMAC)吸附剂,以选择性结合和快速分离His-使用移液器尖端SPE格式标记的蛋白。我们的结果表明,SCF-Ni2 +吸附剂可以选择性地从蛋白质混合物和大肠杆菌细胞裂解物中捕获His标记的蛋白质。综上所述,所开发的方法为纯化带有His标签的蛋白提供了一种快速,便捷和有效的方法。 (C)2015 Elsevier B.V.保留所有权利。

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