Nara Institute of Science and Technology (NAIST), 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
Colloids Surf B Biointerfaces. 2010 Jan 1;75(1):323-9. doi: 10.1016/j.colsurfb.2009.09.004. Epub 2009 Sep 11.
Fixation of ferritin using amino-silane modified substrates is effective, but salt and alkali ions of the buffer can contaminate substrates, inhibiting the sensing and fabrication of nano-electronic devices. To avoid adsorption of salts and alkali ions, buffer solutions have been replaced by pure water or alkali-metal-ion-free buffer. However, proteins in such solutions are sometimes denatured. Therefore, we developed a substrate which adsorbs ferritin but does not adsorb contaminants such as salts and alkali metal-ions contained in the buffer. Adsorption of ferritin was achieved by using a buffer with a high ion strength, such as PBS buffer, because the Debye length becomes shorter with increased ion strength due to intermolecular force even when the substrate has no positive charge. The combination of high coverage methyltrimethoxysilane (MTMS)-coupled silicon substrate and PBS buffer solution is effective for adsorption of ferritin while not adsorbing buffer components such as contaminants and/or salts on the silicon substrate.
使用氨基硅烷修饰的基底固定铁蛋白的方法很有效,但缓冲液中的盐和碱离子会污染基底,从而抑制纳米电子器件的传感和制造。为了避免盐和碱离子的吸附,可以用纯水或无碱金属离子缓冲液替代缓冲溶液。然而,在这样的溶液中的蛋白质有时会变性。因此,我们开发了一种基底,它可以吸附铁蛋白,但不吸附缓冲液中含有的盐和碱金属离子等污染物。由于分子间作用力,随着离子强度的增加,德拜长度会变短,即使基底没有正电荷,也可以使用离子强度较高的缓冲液(如 PBS 缓冲液)来实现铁蛋白的吸附。高覆盖率的甲基三甲氧基硅烷(MTMS)-偶联硅基底与 PBS 缓冲溶液的组合对于铁蛋白的吸附是有效的,而不会在硅基底上吸附缓冲液成分(如污染物和/或盐)。