Blasi L, Longo L, Pompa P P, Manna L, Ciccarella G, Vasapollo G, Cingolani R, Rinaldi R, Rizzello A, Acierno R, Storelli C, Maffia M
National Nanotechnology Laboratory of INFM, c/o Department of Innovation Engineering, University of Lecce, Italy.
Biosens Bioelectron. 2005 Jul 15;21(1):30-40. doi: 10.1016/j.bios.2004.10.012. Epub 2004 Nov 28.
In this paper we have tested two different procedures (the "three-step" and the "four-step" procedures) for the covalent immobilization of glutamate dehydrogenase (GDH) onto silicon supports. Atomic force microscopy (AFM), Fourier-transform infrared spectroscopy (FT-IR), fluorescence spectroscopy and an enzymatic assay were used to probe the structure and activity of the immobilized enzyme. Our results demonstrate that coupling through the "three-step" procedure does not significantly affect either the fold pattern or the activity of the enzyme, suggesting that this method could be ideally suited to the development of high quality monolayers for use in enzyme-based planar biosensors.
在本文中，我们测试了两种不同的将谷氨酸脱氢酶（GDH）共价固定到硅载体上的方法（“三步”法和“四步法”）。利用原子力显微镜（AFM）、傅里叶变换红外光谱（FT-IR）、荧光光谱和酶活性测定来探测固定化酶的结构和活性。我们的结果表明，通过“三步”法进行偶联对酶的折叠模式或活性没有显著影响，这表明该方法可能非常适合用于基于酶的平面生物传感器的高质量单分子层的开发。
