物理化学学报 >> 2017, Vol. 33 >> Issue (4): 810-815.doi: 10.3866/PKU.WHXB201701032

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应用纳米球刻蚀法在自组装膜修饰的硅表面生成中尺度的网状蛋白层

SCHLERETH Andrew,NOOMUNA Panae,GAO Pei*()   

  • 收稿日期:2016-10-31 发布日期:2017-03-23
  • 通讯作者: GAO Pei E-mail:pei.gao@eku.edu
  • 基金资助:
    美国国立卫生研究院(P20GM103436);美国国家科学基金会(3048111570-15-153)

Mesoscale Protein Patterning on a Self-Assembled Monolayer Coated Silicon Surface through Nanosphere Lithography

Andrew SCHLERETH,Panae NOOMUNA,Pei GAO*()   

  • Received:2016-10-31 Published:2017-03-23
  • Contact: Pei GAO E-mail:pei.gao@eku.edu
  • Supported by:
    National Institute of General Medical Sciences of the National Institutes of Health, USA(P20GM103436);National Science Foundation, USA(3048111570-15-153)

摘要:

在自组装膜修饰的硅表面制备有序的蛋白阵列是研发生物传感器的先决条件之一,因此如何产生有序的表面蛋白阵列一直是生物医药研究方向的前沿.本研究通过应用纳米球刻蚀法在氧化的10-烯基十一烷基三氯硅烷自组装膜修饰的硅表面生成了网状结构溶菌酶蛋白层.网孔的大小(从纳米到微米级别)由表面沉积的纳米球的尺寸来调控.我们利用原子力显微镜和荧光显微镜对样品表面进行了详细表征.结果表明:这种新方法比传统的通过扫描探针在固体表面修饰而聚集溶菌酶蛋白的方法更快捷简便,而且它能够在相对大的硅表面形成网状蛋白层.此外,网孔表面附着具有强吸附活性的羧酸基团层,它可以通过静电吸引或者共价结合来吸附液相中的第二种蛋白分子.

关键词: 纳米球刻蚀法, 溶菌酶, 网状阵列, 自组装膜, 原子力显微镜, 荧光显微镜

Abstract:

The patterning and immobilization of protein molecules onto functionalized silicon substrate through surface silane chemistry is of interest because protein patterning is an important prerequisite for the development of protein-based diagnostics in biological and medicinal fields. As a model system, mesoscale netty lysozyme arrays were assembled on oxidized undecyltrichlorosilane (UTSox) monolayer coated silicon surface through nanosphere lithography. The size of the arrays ranged from nanometer to micrometer can be easily adjusted by changing the size of nanospheres applied on the surface. By using nanosphere lithography, we are capable of fabricating a regular array of protein islands over centimeter sample regions. The created lysozyme protein patterns were characterized by atomic force microscopy (AFM) and fluorescence microscope, respectively. The analysis has demonstrated that this newly established approach offers a faster and more reliable process to fabricate netty protein arrays over large areas compared to conventional scanning-probe based fabrication methods. Furthermore, the carboxylic acid-terminated layer on surfaces is particularly effective for immobilizing protein molecules through either electrostatic interactions or covalent attachment via imine bonds. Therefore, the negative-toned protein structure on the surface with carboxylic acid groups coated on the bare areas makes it possible to fabricate two types of protein molecules on one surface.

Key words: Nanosphere lithography, Lysozyme, Netty array, Self-assembled monolayer, Atomic force microscopy, Fluorescence microscope

MSC2000: 

  • O647