Submicron streptavidin patterns for protein assembly
Submicron streptavidin patterns for protein assembly
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DOI：
10.1021/la0608213
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发表时间：
2006-08-15
期刊：
影响因子：
3.9
通讯作者：
Maynard, Heather D.
中科院分区：
文献类型：
--
作者：
Christman, Karen L.;Requa, Michael V.;Maynard, Heather D.
利用光刻技术在聚合物薄膜中制备微米和亚微米尺度的醛官能团，以开发生物相关尺度的蛋白质固定化和组装平台。含有pH反应性聚合物聚(3，3‘-甲基丙烯酸二乙氧基丙酯)和光酸产生器(PAG)的薄膜通过365 nm(I线)光照射从500 nm到40微米被图案化。在PAG活化和缩醛的水解后，形成了醛基团。用生物素化的乙醛反应探针孵育薄膜后，X射线光电子能谱结果与生物素附着在薄膜表面的结果一致。背景随后通过洪水暴露和与端氨基氧基的聚乙二醇孵育而钝化，导致非特异性蛋白质吸附减少98%。用链霉亲和素、生物素标记的炭疽毒素受体1和炭疽毒素的保护性抗原部分证明了蛋白质的构型和组装，并用荧光显微镜和原子力显微镜(AFM)进行了证实。AFM显示获得了500 nm的蛋白质特征。由于生物素化蛋白质的丰富，这种方法为蛋白质的固定化和组装提供了一个平台，在生物技术中的各种应用。
Micron and submicron-scale features of aldehyde functionality were fabricated in polymer films by photolithography to develop a platform for protein immobilization and assembly at a biologically relevant scale. Films containing the pH-reactive polymer poly(3,3'-diethoxypropyl methacrylate) and a photoacid generator (PAG) were patterned from 500 nm to 40 mu m by exposure to 365 nm (i-line) light. Upon PAG activation and hydrolysis of acetals, aldehyde groups formed. After the films were incubated with a biotinylated aldehyde reactive probe, the X-ray photoelectron spectroscopy results were consistent with biotin being attached to the surface. The background was subsequently passivated by flood exposure and incubation with an aminooxy-terminated poly(ethylene glycol), resulting in a 98% reduction in nonspecific protein adsorption. Protein patterning and assembly was demonstrated using streptavidin, biotinylated anthrax toxin receptor-1, and the protective antigen moiety of anthrax toxin and confirmed by fluorescence microscopy and atomic force microscopy (AFM). AFM demonstrated that 500 nm protein features were achieved. Because of the abundance of biotinylated proteins, this methodology provides a platform for protein immobilization and assembly for various applications in biotechnology.