A novel peptide microarray for protein detection and analysis utilizing a dry peptide array system
A novel peptide microarray for protein detection and analysis utilizing a dry peptide array system
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DOI：
10.1039/b514263f
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发表时间：
2006-02-01
影响因子：
--
通讯作者：
Mihara, H
中科院分区：
文献类型：
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作者：
Usui, K;Tomizaki, K;Mihara, H
构建了一种新型干肽微阵列系统，为蛋白质检测和分析提供了一种实用的解决方案。该系统是在干燥条件下的阵列制备和分析程序，使用设计的肽作为非固定化捕获剂来检测蛋白质。该系统有几个优点，包括其便携性和易用性，以及不需要考虑样品溶液蒸发的事实。在这项研究中，各种蛋白质已经用a-螺旋肽迷你文库进行了表征。当蛋白质加入到肽库阵列时，荧光肽根据其序列表现出不同的荧光强度。这些反应的模式可以被视为蛋白质指纹图谱，这足以确定目标蛋白的身份。此外，使用聚类分析对所得PFPs进行统计分析。根据蛋白的家族和结合特性，成功地聚集了蛋白的pfp。此外，目标蛋白使用纳升系统进行表征，可以检测到低至1.2 fmol。这些研究表明，干肽阵列系统是一种很有前途的检测和分析目标蛋白的方法。干肽阵列将在高通量蛋白质检测纳米/微阵列的发展中发挥重要作用，用于蛋白质组学和配体筛选研究。
A novel dry peptide microarray system has been constructed that affords a practical solution for protein detection and analysis. This system is an array preparation and assay procedure tinder dry conditions that uses designed peptides as non-immobilized capture agents for the detection of proteins. The system has several advantages that include its portability and ease-Of-Use, as well as the fact that vaporization of sample solutions need not be considered. In this study, various proteins have been characterized with an a-helical peptide mini-library. When proteins were added to the peptide library array, the fluorescent peptides showed different fluorescent intensities depending on their sequences. The patterns of these responses could be regarded as protein fingerprints' (PFPs), which are sufficient to establish the identities of the target proteins. Furthermore, statistical analysis of the resulting PFPs was performed using cluster analysis. The PFPs of the proteins were clustered successfully depending on their families and binding properties. Additionally, the target protein was characterized using a nanolitre system and could be detected down to 1.2 fmol. These Studies imply that the dry peptide array system is a promising too] for detecting and analyzing target proteins. The dry peptide array will play a role in development of high-throughput protein-detecting nano/micro arrays for proteomics and ligand screening studies.