Horner Scott R, Mace Charles R, Rothberg Lewis J, Miller Benjamin L
Department of Biochemistry and Biophysics, University of Rochester, NY 14642, USA.
Biosens Bioelectron. 2006 Feb 15;21(8):1659-63. doi: 10.1016/j.bios.2005.07.019. Epub 2005 Sep 8.
The study of proteins and the molecules with which they interact on an organismwide scale is critical to understanding basic biology, and understanding and improving human health. New platform technologies allowing label-free, quantitative array-based analysis of proteins are particularly desirable. We have developed an analytical technology, reflective interferometry (RI), which provides specific, rapid, and label-free optical detection of biomolecules in complex mixtures. In order to evaluate the suitability of RI for proteomics, we have prepared a series of arrays bearing the extracellular domain of the secreted enteropathogenic Escherichia coli (EPEC) protein Translocated Intimin Receptor (Tir). These arrays are able to selectively detect the extracellular domain of the protein Intimin, Tir's natural binding partner. Furthermore, we demonstrate the use of RI and Tir-functionalized arrays for the selective detection of EPEC directly from culture.
在全生物体范围内研究蛋白质及其相互作用的分子对于理解基础生物学以及了解和改善人类健康至关重要。尤其需要能够对蛋白质进行无标记、基于阵列的定量分析的新平台技术。我们开发了一种分析技术——反射干涉测量法（RI），它能够对复杂混合物中的生物分子进行特异性、快速且无标记的光学检测。为了评估RI在蛋白质组学中的适用性，我们制备了一系列带有分泌型肠致病性大肠杆菌（EPEC）蛋白转位紧密素受体（Tir）胞外结构域的阵列。这些阵列能够选择性地检测紧密素（Tir的天然结合伴侣）的胞外结构域。此外，我们展示了使用RI和Tir功能化阵列直接从培养物中选择性检测EPEC的方法。
