Stable Microstructured Network for Protein Patterning on a Plastic Microfluidic Channel: Strategy and Characterization of On-Chip Enzyme Microreactors
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- 作者：Haiyun Qu ; Haitao Wang ; Yi Huang ; Wei Zhong ; Haojie Lu ; Jilie Kong ; Pengyuan Yang ; and Baohong Liu
- 刊名：Analytical Chemistry
- 出版年：2004
- 出版时间：November 1, 2004
- 年：2004
- 卷：76
- 期：21
- 页码：6426 - 6433
- 全文大小：148K
- 年卷期：v.76,no.21(November 1, 2004)
- ISSN：1520-6882
文摘
Chemical modification of a poly(methyl methacrylate)(PMMA) microchannel surface has been explored tofunctionalize microfluidic chip systems. A craft copolymerwas designed and synthesized to introduce the silanefunctional groups onto the plastic surface first. Furthermore, it has been found that, through a silicon-oxygen-silicon bridge that formed by tethering to these functionalgroups, a stable patterning network of gel matrix couldbe achieved. Thus, anchorage of proteins could be realizedonto the hydrophobic PMMA microchannels with bioactivity preserved as far as possible. The protein homogeneous patterning in a microfluidic channel has beendemonstrated by performing microchip capillary electrophoresis with laser-induced fluorescence detection andconfocal fluorescence microscopy. To investigate thebioactivity of enzymes entrapped within stable silica gel-derived microchannels, the suggested scheme was employed to the construction of immobilized enzyme microreactor-on-a-chip. The proteolytic activity of immobilizedtrypsin has been demonstrated with the digestion ofcytochrome c and bovine serum albumin at a fast flow rateof 4.0 
L/min, which affords the short residence time lessthan 5 s. The digestion products were characterized usingMALDI-TOF MS with sequence coverage of 75 and 31%observed, respectively. This research exhibited a simplebut effective strategy of plastic microchip surface modification for protein immobilization in biological and proteomic research.
L/min, which affords the short residence time lessthan 5 s. The digestion products were characterized usingMALDI-TOF MS with sequence coverage of 75 and 31%observed, respectively. This research exhibited a simplebut effective strategy of plastic microchip surface modification for protein immobilization in biological and proteomic research.