Multiplexed Lipid Dip-Pen Nanolithography on Subcellular Scales for the Templating of Functional Proteins and Cell Culture
Multiplexed Lipid Dip-Pen Nanolithography on Subcellular Scales for the Templating of Functional Proteins and Cell Culture
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DOI：
10.1002/smll.200800949
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发表时间：
2008-10-01
期刊：
影响因子：
13.3
通讯作者：
Lenhert, Steven
中科院分区：
文献类型：
--
作者：
Sekula, Sylwia;Fuchs, Jeanette;Lenhert, Steven
生物系统中发生的分子图案化过程由于其动态行为、亚细胞尺寸和高度复杂性而在体内研究具有挑战性。使用纳米光刻技术对生物分子进行体外图案化可以简化过程并详细研究动态相互作用。平行蘸笔纳米光刻（DPN）由于其结构性、高分辨率和高通量而独特地能够在亚细胞长度尺度上整合功能性生物分子。磷脂特别适合作为DPN的油墨，因为各种不同的功能性脂质可以容易地平行图案化。在此，DPN用于空间图案化多组分微米和纳米结构的支撑脂质膜和多层，其为流体并且含有各种量的生物素和/或次氮基三乙酸官能团。用荧光显微镜和光电子发射显微镜对图案进行了表征。基于链霉亲和素或组氨酸-标签偶联的功能化或重组蛋白的选择性吸附使得能够半合成制造模型外周膜结合蛋白。然后将以这种方式形成的仿生膜图案用作细胞培养的基底，如通过T细胞的选择性粘附和活化所证明的。
Molecular patterning processes taking place in biological systems are challenging to study in vivo because of their dynamic behavior, subcellular size, and high degree of complexity. In vitro patterning of biomolecules using nanolithography allows simplification of the processes and detailed study of the dynamic interactions. Parallel dip-pen nanolithography (DPN) is uniquely capable of integrating functional biomolecules on subcellular length scales due to its constructive nature, high resolution, and high throughput. Phospholipids are particularly well suited as inks for DPN since a variety of different functional lipids can be readily patterned in parallel. Here DPN is used to spatially pattern multicomponent micro- and nano-structured supported lipid membranes and multilayers that are fluid and contain various amounts of biotin and/or nitrilotriacetic acid functional groups. The patterns are characterized by fluorescence microscopy and photoemission electron microscopy. Selective adsorption of functionalized or recombinant proteins based on streptavidin or histidine-tag coupling enables the semisynthetic fabrication of model peripheral membrane bound proteins. The biomimetic membrane patterns formed in this way are then used as substrates for cell culture, as demonstrated by the selective adhesion and activation of T-cells.