Patterning ligands on reactive SAMs by microcontact printing
Patterning ligands on reactive SAMs by microcontact printing
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DOI：
10.1021/la9811970
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发表时间：
1999-03-16
期刊：
影响因子：
3.9
通讯作者：
Whitesides, GM
中科院分区：
文献类型：
--
作者：
Lahiri, J;Ostuni, E;Whitesides, GM
本文描述了一种用微接触印刷（mu CP）在金上的烷硫酸酯混合SAMs上印制配体图案的方法。混合SAMs由末端有三（乙二醇）基团(HS(CH2)(11)（OCH2CH2)(3)OH, 1）和末端有六（乙二醇）-CH2CO2H基团(HS(CH2)(11)（OCH2CH2)(6)OCH2CO2H, 2）的硫醇组成。配体的打印过程分为两步。2的羧基首先转化为活性的五氟苯基酯。然后将新氧化的PDMS图章与伯胺衍生的配体连接，与活化的SAM接触；在接触区域，胺与被激活的胺发生反应。酯和形成酰胺。两种配体，生物素和苯磺酰胺，被印在这些sam上。将底物与荧光标记的抗菌素抗体溶液孵育，通过荧光显微镜检测呈递生物素配体的图案sam的形成。利用夹心实验也检测了图案生物素的形成；在本实验中，SAM在链霉亲和素、蛋白g -生物素偶联物和荧光标记的山羊抗兔IgG溶液中依次孵育。这些方法得到的图像中最小的特征是边长为5 μ m的正方形。利用表面等离子体共振（SPR）检测抗生素素抗体与具有生物素基团的SAMs的结合，估计mu CP的偶联率与浸泡法的偶联率接近90%。通过碳酸酐酶（CA）与磺胺衍生物sam的结合来检测苯磺酰胺配体的打印；根据SPR估计，偶联的产率与浸泡法的75%相似。对于这两种配体，发现在油墨之前氧化PDMS印记对良好的偶联产率至关重要。
This report describes a method for patterning ligands onto mixed SAMs of alkanethiolates on gold by microcontact printing (mu CP). The mixed SAMs were made from thiols presenting terminal tri(ethylene glycol) groups (HS(CH2)(11)(OCH2CH2)(3)OH, 1) and terminal hexa(ethylene glycol)-CH2CO2H groups (HS(CH2)(11)(OCH2CH2)(6)OCH2CO2H, 2). Ligands were printed using a two-step procedure. The carboxylic acid groups of 2 were first converted to reactive pentafluorophenyl esters. A freshly oxidized PDMS stamp, inked with a ligand derivatized with a primary amine, was then brought into contact with the activated SAM; in the areas of contact, the amine reacted with the activated. ester and formed an amide. Two ligands, biotin and benzenesulfonamide, were printed onto these SAMs. The formation of patterned SAMs presenting biotin ligands was detected by fluorescence microscopy of substrates that were incubated with a solution of fluorescently labeled antibiotin antibody. The formation of patterned biotin was also detected using a sandwich experiment; in this experiment, the SAM was incubated sequentially in solutions of streptavidin, protein G-biotin conjugate, and fluorescently labeled goat antirabbit IgG. The smallest features resolved in images obtained by these methods were squares with a 5 mu um side. Using surface plasmon resonance (SPR) to detect binding of antibiotin antibody to SAMs presenting biotin groups, the yield of coupling by mu CP was estimated to be similar to 90% of that obtained by immersion. Printing of the benzenesulfonamide ligand was detected by binding of carbonic anhydrase (CA) to the sulfonamide-derivatized SAMs; the yield of coupling, as estimated by SPR, was similar to 75% of that obtained by immersion. For both ligands, oxidation of the PDMS stamp before inking was found to be critical for good coupling yields.