Functionalization of poly(oligo(ethylene glycol)methacrylate) films on gold and Si/SiO2 for immobilization of proteins and cells:: SPR and QCM studies
Functionalization of poly(oligo(ethylene glycol)methacrylate) films on gold and Si/SiO2 for immobilization of proteins and cells:: SPR and QCM studies
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DOI：
10.1021/bm7009043
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发表时间：
2007-12-01
影响因子：
6.2
通讯作者：
Choi, Insung S.
中科院分区：
文献类型：
--
作者：
Lee, Bong Soo;Chi, Young Shik;Choi, Insung S.
利用原子转移自由基聚合(ATRP)引发剂溴酸酯为引发剂的自组装单分子膜(SAM)的形成，在金和Si/SiO_2衬底上制备了不同厚度的生物相容性和无生物污染的聚甲基丙烯酸寡乙醇酯(POEGMA)薄膜。在pOEGMA膜形成后，用N，N‘-二琥珀酰亚胺碳酸酯(DSC)活化甲基丙烯酸酯骨架上的侧链末端羟基，然后将DSC活化的pOEGMA膜与(+)-生物素-3，6，9-三氧杂十二胺(Biotin-NH2)反应生成生物素化的pOEGMA膜。通过与目标蛋白(链霉亲和素)和模型蛋白(纤维蛋白原和溶菌酶)的表面等离子体共振实验，我们证实了所得到的膜与链霉亲和素的生物特异性结合的信噪比比由羧酸封端的自组装膜制备的生物素化底物提高了10倍。用石英晶体微天平测量了链霉亲和素和纤维蛋白原在不同厚度的生物素化pOEGMA膜上的表面覆盖率，并考察了膜厚度对生物特异性结合的影响。链霉亲和素。在20 nm厚的POEGMA膜上，链霉亲和素的结合量和链霉亲和素/纤维蛋白原的信噪比均达到饱和。此外，为了展示pOEGMA膜的广泛适用性，我们分别在DSC活化的pOEGMA膜上通过微接触印刷生物素-NH_2和聚L-赖氨酸构建了链霉亲和素和细胞的微图案。
Thin films of a biocompatible and nonbiofouling poly(oligo(ethylene glycol) methacrylate) (pOEGMA) with various thicknesses were formed on gold and Si/SiO2 substrates by a combination of the formation of self-assembled monolayers (SAMs) terminating in bromoester-an initiator of atom transfer radical polymerization (ATRP)-and surface-initiated ATRP. After the formation of the pOEGMA films, terminal hydroxyl groups of side chains divergent from the methacrylate backbones were activated with N,N'-disuccinimidyl carbonate (DSC), and the DSC-activated pOEGMA films were reacted with (+)-biotinyl-3,6,9-trioxatmdecanediamine (Biotin-NH2) to form biotinylated pOEGMA films. By surface plasmon resonance experiments with the target protein (streptavidin) and model proteins (fibrinogen and lysozyme), we verified that the resulting films showed the enhanced signal-to-noise ratio (similar to 10-fold enhancement) for the biospecific binding of streptavidin compared with the biotinylated substrate prepared from carboxylic acid-terminated SAMs. Quartz crystal microbalance measurements were also carried out to obtain the surface coverage of streptavidin and fibrinogen adsorbed onto the biotinylated pOEGMA films with various thicknesses and to investigate the effect of film thicknesses on the biospecific binding of. streptavidin. Both the binding capacity of streptavidin and the signal-to-noise ratio of streptavidin/fibrinogen were found to be saturated at the 20 nm thick pOEGMA film. In addition, to demonstrate a wide applicability of the pOEGMA films, we constructed micropatterns of streptavidin and cells by microcontact-printing biotin-NH2 and poly-L-lysine onto the DSC-activated pOEGMA films, respectively.