The determination of thickness and surface mass density of mesothick immunoprecipitate layers by null ellipsometry and protein 125iodine labeling
The determination of thickness and surface mass density of mesothick immunoprecipitate layers by null ellipsometry and protein 125iodine labeling
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DOI：
10.1006/jcis.2002.8247
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发表时间：
2002-05-01
影响因子：
9.9
通讯作者：
Tengvall, P
中科院分区：
文献类型：
--
作者：
Benesch, J;Askendal, A;Tengvall, P
本研究的目的是用椭圆偏振测量法在疏水硅表面交替沉积HSA和多克隆抗HSA层，以确定厚达110 nm的有机层的厚度和表面质量密度。将椭圆偏振法测得的厚度与原子力显微镜测得的厚度进行比较，并用I-125标记的蛋白质校准沉积表面质量密度。结果表明，当蛋白质膜折射率N-膜=1.5-0I时，原子力显微镜和椭偏仪测量的蛋白质层厚度有很好的一致性，而根据椭偏仪数据计算的表面质量密度被Cuypers单组分公式严重高估。当公式中的M/A比从4.14降至2.35时，表面质量密度得到了很好的吻合。这种方法为测定固体载体上的中间有机层蛋白质的折射率和表面质量密度提供了一种方便的方法。(C)2002年埃尔塞维尔科学公司(美国)。
The aim of the present study was to ellipsometrically determine the thickness and surface mass density in air for up to 110-nm-thick organic layers made of alternatingly deposited layers of HSA and polyclonal anti-HSA on hydrophobic silicon. The ellipsometrically determined thickness was compared to that obtained by AFM and the deposited surface mass density calibrated with I-125-labeled proteins. The results indicate a good agreement in protein layer thickness between AFM and ellipsometry when the protein film refractive index N-film = 1.5 - 0i, although then the calculated surface mass density from the ellipsometry data became grossly overestimated by the Cuypers one-component formula. A good agreement in the surface mass density was obtained when the M/A ratio in this formula was lowered from 4.14 to 2.35. This approach indicates a convenient means of determining the refractive indices and surface mass densities of mesothick organic layers proteins on solid Supports. (C) 2002 Elsevier Science (USA).