Immobilization of histidine-tagged recombinant proteins onto micropatterned surfaces for cell-based functional assays
Immobilization of histidine-tagged recombinant proteins onto micropatterned surfaces for cell-based functional assays
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DOI：
10.1021/la050893e
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发表时间：
2005-08-02
期刊：
影响因子：
3.9
通讯作者：
Iwata, H
中科院分区：
文献类型：
--
作者：
Kato, K;Sato, H;Iwata, H
这封信描述了一种制备蛋白质微阵列的方法，该方法允许在细胞水平上对蛋白质进行功能分析。该方法涉及利用经遗传工程改造以携带对金属离子具有亲和力的融合标签的重组蛋白。使用微图案化的烷醇单分子层来制备具有固定金属离子的多个斑点的微阵列。在生理条件下，融合蛋白被螯合到斑点上。通过在显示寡组氨酸标记的表皮生长因子的微阵列上培养神经干细胞，证明了该方法的可行性。
This letter describes a method for preparing protein microarrays that allow the functional analysis of proteins at a cellular level. This method involves the utilization of recombinant proteins genetically engineered to carry a fusion tag that has an affinity for metal ions. A micropatterned alkanethiol monolayer was used to prepare a microarray having multiple spots with immobilized metal ions. The fusion protein was chelated to the spots under physiological conditions. The feasibility of the method was demonstrated by culturing neural stem cells on the microarray that displayed oligohistidine-tagged epidermal growth factor.