Single-molecule selection and recovery of structure-specific antibodies using atomic force microscopy
Single-molecule selection and recovery of structure-specific antibodies using atomic force microscopy
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DOI：
10.1016/j.nano.2007.06.001
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发表时间：
2007-09-01
影响因子：
5.4
通讯作者：
Sierks, Michael R.
中科院分区：
文献类型：
--
作者：
Shlyakhtenko, Luda S.;Yuan, Bin;Sierks, Michael R.
蛋白质错误折叠和聚集是包括阿尔茨海默氏病、帕金森氏病、亨廷顿氏病、肌萎缩性侧索硬化症、糖尿病和朊病毒相关疾病在内的许多疾病中的共同线索。阐明各种蛋白质形式在这些疾病中所起的作用需要能够靶向特定蛋白质形式的试剂。在这里，我们提出了一种方法来分离抗体结合到一个特定的蛋白质形式。我们将原子力显微镜（AFM）的成像和纳米操纵能力与噬菌体展示抗体库的蛋白质多样性相结合，开发了一种技术，使我们能够回收与单个蛋白质分子靶点结合的单个抗体分子。首先通过AFM对靶蛋白-抗体复合物进行成像，然后通过纳米光刻法在靶抗体上操作AFM针尖以回收相关的噬菌体，并通过聚合酶链反应从单个噬菌体颗粒中回收抗体基因。(C)2007年由Elsevier Inc.出版
Protein misfolding and aggregation are a common thread in numerous diseases including Alzheimer's, Parkinson's, Huntington's, amyotrophic lateral sclerosis, diabetes, and prion-related diseases. Elucidation of the role played by the various protein forms in these diseases requires reagents that can target specific protein forms. Here we present a method to isolate antibodies that bind to a specific protein form. We combined the imaging and nanomanipulation capabilities of atomic force microscopy (AFM) with the protein diversity of phage display antibody libraries to develop a technology that allows us to recover a single antibody molecule that is bound to a single protein molecular target. The target protein-antibody complex is first imaged by AFM, the AFM tip is then manipulated by nanolithography over the target antibody to recover the associated phage, and the antibody gene is recovered from the single phage particle by polymerase chain reaction. (C) 2007 Published by Elsevier Inc.