A general method for patterning gradients of biomolecules on surfaces using microfluidic networks
A general method for patterning gradients of biomolecules on surfaces using microfluidic networks
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DOI：
10.1021/ac048440m
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发表时间：
2005-04-15
影响因子：
7.4
通讯作者：
Whitesides, GM
中科院分区：
文献类型：
--
作者：
Jiang, XY;Xu, QB;Whitesides, GM
Ibis报告概述了在表面上制造生物分子固定梯度的一般方法。该方法利用微流体网络，其在溶液中产生抗生物素蛋白的梯度，并通过物理吸附将该蛋白质固定在玻璃或聚（二甲基硅氧烷）的表面上。然后将抗生物素蛋白的固定梯度翻译成生物素化配体（例如，小分子、DNA低聚物、多糖），使用生物素和抗生物素蛋白之间的特异性相互作用。该方法还可以通过将梯度从溶液直接转移到表面上来产生某些蛋白质和人工聚合物的固定化梯度。这种方法的主要优点是，原则上，几乎任何类型的分子都可以固定在任意形状的定义明确的表面梯度中，尺寸为几微米到几厘米。可以从溶液中的梯度中调整表面上梯度的精确形状，无论是动力学上还是竞争性地。动力学方法依赖于控制表面暴露于溶液中的梯度的时间：当单个蛋白质从溶液中吸附时，吸附的量取决于其在溶液中的浓度和允许吸附的时间。竞争性方法依赖于将表面暴露于溶液中两种蛋白质的互补梯度（在这些实验中，溶液中蛋白质浓度的总和与位置无关，尽管每种蛋白质的浓度单独取决于位置。在这个过程中，每种蛋白质在表面饱和时的相对量仅取决于其浓度）。
Ibis report outlines a general method for the fabrication of immobilized gradients of biomolecules on surfaces. This method utilizes a microfluidic network that generates a gradient of avidin in solution and immobilizes this protein on the surface of glass or poly(dimethylsiloxane) by physical adsorption. The immobilized gradient of avidin is then translated into gradients of biotinylated ligands (e.g., small molecules, oligomers of DNA, polysaccharides) using the specific interaction between biotin and avidin. This method can also generate immobilized gradients of certain proteins and artificial polymers by a direct transfer of gradients from solution onto the surface. The major advantage of this method is that almost any type of molecule can, in principle, be immobilized in a well-defined surface gradient of arbitrary shape with dimensions of a few micrometers to a few centimeters. It is possible to tailor the precise shapes of gradients on surfaces from gradients in solution, either kinetically or competitively. Kinetic methods rely on controlling the time that the surface is exposed to the gradient in solution: when a single protein adsorbs from solution, the amount that adsorbs depends both on its concentration in solution and on the time allowed for adsorption. Competitive methods rely on exposure of the surface to a complementary gradient of two proteins in solution (In these experiments, the sum of the concentrations of the proteins in solution is independent of positions although the concentration of each, individually, depends on the position. In this procedure, the relative amount of each protein, at saturation on the surface, depends only on its concentration).