Individually addressable parallel peptide synthesis on microchips
Individually addressable parallel peptide synthesis on microchips
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DOI：
10.1038/nbt723
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发表时间：
2002-09-01
影响因子：
46.9
通讯作者：
Gao, XL
中科院分区：
文献类型：
--
作者：
Pellois, JP;Zhou, XC;Gao, XL
小型化的、空间可寻址的肽和肽模拟物微芯片是高通量生物医学和药物研究以及蛋白质组学进步的有力工具。在这里，我们报告了一种有效的和灵活的方法，用于在可单独寻址的微芯片上并行合成肽，在脱保护步骤中使用数字光刻和光生酸。我们证明，我们能够使用20种天然氨基酸以及合成氨基酸类似物在微芯片上1平方厘米的区域内合成数千种肽，并且分步产率高，反应周期时间短。使用p53抗体（PAb240）的表位筛选实验产生明确定义的结合模式。微芯片上的拟肽序列显示特异性抗体结合，并提供对负责表位结合特异性的分子细节的了解。我们的方法只需要一个传统的合成器和一个计算机控制的光学模块，从而允许在常规研究实验室的各种药物和蛋白质组学应用的肽微芯片的潜在发展。
Miniaturized, spatially addressable microchips of peptides and peptidomimetics are powerful tools for high-throughput biomedical and pharmaceutical research and the advancement of proteomics. Here we report an efficient and flexible method for the parallel synthesis of peptides on individually addressable microchips, using digital photolithography and photogenerated acid in the deprotection step. We demonstrate that we are able to synthesize thousands of peptides in a 1 cm 2 area on a microchip using 20 natural amino acids as well as synthetic amino acid analogs, with high stepwise yields and short reaction-cycle times. Epitope screening experiments using a p53 antibody (PAb240) produced clearly defined binding patterns. The peptidomimetic sequences on the microchip show specific antibody binding and provide insights into the molecular details responsible for specificity of epitope binding. Our approach requires just a conventional synthesizer and a computer-controllable optical module, thereby allowing potential development of peptide microchips for various pharmaceutical and proteomic applications in routine research laboratories.