Suspension arrays of hydrogel microparticles prepared by photopatterning for multiplexed protein-based bioassays
Suspension arrays of hydrogel microparticles prepared by photopatterning for multiplexed protein-based bioassays
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DOI：
10.1007/s10544-008-9196-1
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发表时间：
2008-12-01
影响因子：
2.8
通讯作者：
Koh, Won-Gun
中科院分区：
文献类型：
--
作者：
Lee, Woojin;Choi, Dongkil;Koh, Won-Gun
已经使用形状编码的聚（乙二醇）（PEG）水凝胶微粒开发了用于基于蛋白质的测定的悬浮阵列，以克服使用颜色编码的刚性微粒作为蛋白质支持物的当前系统的问题。采用聚二甲基硅氧烷（PDMS）通道作为成型嵌件，通过两步法制备了各种形状的水凝胶微粒，该方法包括预定型和冲洗。使用不同分子量的PEG（700、3，400和8，000 Da）制造横向尺寸在50至300 μ m范围内的水凝胶微粒，由此可以控制水凝胶微粒的水含量和溶胀行为。以悬浮阵列形式制备了蛋白质包埋的水凝胶微粒，由于PEG水凝胶的高含水量和柔软性，其可以包封蛋白质而不失活一周。利用荧光检测技术成功地研究了水凝胶包埋的葡萄糖氧化酶（GOX）和过氧化物酶（POD）的顺序双酶反应，展示了悬浮阵列的一种可能的应用。此外，分别含有GOX/POD和碱性磷酸酶（AP）的两种不同形状的水凝胶微粒的混合物的制备和形状编码的悬浮阵列用于同时表征两种不同的酶催化反应。
Suspension arrays for protein-based assays have been developed using shape-coded poly(ethylene glycol) (PEG) hydrogel microparticles to overcome the problems with current systems which use color-coded rigid microparticles as protein supports. Various shapes of hydrogel microparticles were fabricated by a two-step process consisting of photopatterning and flushing using a poly(dimethylsiloxane) (PDMS) channel as a molding insert. Hydrogel microparticles with lateral dimensions ranging from 50 to 300 mu m were fabricated using different molecular weights of PEG (700, 3,400, and 8,000 Da), by which the water content and swelling behavior of the hydrogel microparticles could be controlled. Protein-entrapped hydrogel microparticles were prepared in a suspension array format, and PEG hydrogel could encapsulate proteins without deactivation for a week due to its high water content and soft nature. The sequential bienzymatic reaction of hydrogel-entrapped glucose oxidase (GOX) and peroxidase (POD) was successfully investigated using fluorescence detection, demonstrating one possible application of suspension arrays. Furthermore, a mixture of two different shapes of hydrogel microparticles containing GOX/POD and alkaline phosphatase (AP), respectively, was prepared and the shape-coded suspension array was used for simultaneous characterization of two different enzyme-catalyzed reactions.