Department of Chemical Engineering, University of California, Santa Barbara, CA 93106, USA.
Proc Natl Acad Sci U S A. 2010 Aug 17;107(33):14627-32. doi: 10.1073/pnas.1004302107. Epub 2010 Aug 2.
Direct determination of functional biomolecular chemistry of clinically relevant tissues in vivo is a challenging task. Current approaches, based on tissue retrieval by biopsy and subsequent solubilization, are limited in terms of accurate representation of tissue constituents, reproducibility, and retention of functionality of solubilized tissue biomolecules. Using a pool of known surfactants, we designed and screened a large combinatorial library of surfactant formulations, which led to the discovery of rare synergistic formulations that greatly enhance tissue solubilization as well as preserve bioactivity of solubilized molecules, in particular proteins. By combining these formulations with a short ultrasound application, we developed a tissue sampling method--STAMP (Surfactant-based Tissue Acquisition for Molecular Profiling)--for rapid one-step determination of functional tissue chemistry in vivo. We specifically demonstrate STAMP-assisted profiling of a multitude of proteins, lipids, and genomic DNA in skin and mucosal tissues. Applications of this sampling methodology to rapid molecular diagnostics of cutaneous allergies and infectious diseases are also presented.
在体直接测定临床相关组织的功能性生物分子化学是一项具有挑战性的任务。目前基于活检组织采集和随后的溶解的方法，在组织成分的准确代表性、重现性和溶解组织生物分子的功能保留方面受到限制。我们使用一组已知的表面活性剂，设计和筛选了一个大型的表面活性剂配方组合文库，发现了一些罕见的协同配方，这些配方大大增强了组织的溶解能力，同时保持了溶解分子（特别是蛋白质）的生物活性。通过将这些配方与短时间的超声应用相结合，我们开发了一种组织采样方法——STAMP（基于表面活性剂的分子分析组织采集），用于快速一步法在体测定功能性组织化学。我们特别证明了 STAMP 辅助分析皮肤和粘膜组织中的多种蛋白质、脂质和基因组 DNA。还介绍了这种采样方法在皮肤过敏和传染病的快速分子诊断中的应用。
