Fabrication of dense, uniform aminosilane monolayers: A platform for protein or ligand immobilization
Fabrication of dense, uniform aminosilane monolayers: A platform for protein or ligand immobilization
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DOI：
10.1016/j.colsurfa.2012.08.049
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发表时间：
2012-11-20
影响因子：
5.2
通讯作者：
Subramanian, Anuradha
中科院分区：
文献类型：
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作者：
Thakurta, Sanjukta Guha;Subramanian, Anuradha
设计具有增强的生物分子结合能力的生物界面，以及它们在表面的分布和呈现，在很大程度上需要控制底层的表面化学。为了将实验参数与表面性质、配体分布和生物分子结合联系起来，测定了(3-氨丙基)三乙氧基硅烷(APTES)浓度(1，2和4%)和硅烷化反应时间(5-60min)对可用氨基数目的影响，并与表面结合的人免疫球蛋白G(HIGG)的量进行了关联。用2Vol%APTES硅烷化30min的表面产生致密的硅烷单分子层(1.0-1.2 nm)，其中平均分子取向相对于表面法线为+/-2度。获得了接近10(15)/cm(2)的-NH2部分的表面密度，与所评价的其他条件相比明显更高。椭圆偏振测量和原子力显微镜(AFM)分析的综合数据支持了我们的发现。用2%APTES硅烷化30分钟后，HIGG的偶联率接近10(13)Ig/cm(2)，显著高于1%APTES作用30分钟所得的结果。当用纳米金(R)抗体结合物显示时，固定化的HIGG在用2Vol%APTES硅烷化的表面上分布致密和均匀。这项研究强调了影响功能化表面生物分子固定过程的实验参数的关键作用。爱思唯尔出版公司(Elsevier B.V.)
Designing biointerfaces with enhanced biomolecule binding abilities, along with their distribution and presentation on the surface, largely requires control over underlying surface chemistries. In an effort to correlate experimental parameters to surface properties, ligand distribution and biomolecule binding, the influence of (3-aminopropyl)triethoxysilane (APTES) concentration (1,2 and 4 vol%) and duration of the silanization reaction (5-60 min) on the number of available -NH2 groups were determined and correlated with the amount of surface bound human immunoglobulin G (HIgG). Surfaces silanized with 2 vol% APTES for 30 min yielded a densely populated silane monolayer (1.0-1.2 nm) where the average molecular orientation was 38+/-2 degrees with respect to the surface normal. A surface density of -NH2 moieties of similar to 10(15)/cm(2) was obtained and was significantly higher compared to other conditions evaluated. The combined data from ellipsometry and atomic force microscopy (AFM) analyses supports our findings. Coupling of HIgG to surfaces silanized with 2 vol% APTES for 30 min yielded similar to 10(13) IgG/cm(2), which was significantly higher than values obtained at 1 vol% APTES for 30 min. The distribution of immobilized HIgG was noted to be dense and uniform on surfaces silanized with 2 vol% APTES when visualized using Nanogold(R) antibody conjugates. This study highlights the critical role of the experimental parameters that impact the biomolecule immobilization process on functionalized surfaces. Published by Elsevier B.V.