| 基于渗透脱水的自动化蛋白质结晶高通量筛选芯片 |
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| 引用本文： | 罗娅慧,李刚,陈强,赵建龙.基于渗透脱水的自动化蛋白质结晶高通量筛选芯片[J].高等学校化学学报,2012,33(10):2178-2183. |
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| 作者姓名： | 罗娅慧 李刚 陈强 赵建龙 |
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| 作者单位： | 1. 中国科学院上海微系统与信息技术研究所, 传感器技术联合国家重点实验室, 上海 200050;
2. 中国科学院研究生院, 北京 100049 |
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| 基金项目： | 国家"九七三"计划项目，国家自然科学基金，中国科学院科研装备研制项目，上海市自然科学基金 |
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| 摘 要： | 构建了一种基于渗透脱水模式的自动进样微流控结晶芯片. 该芯片通过真空预脱气将包含蛋白质和结晶剂的液滴自动分配至结晶微腔阵列中, 然后利用集成的一排包含不同浓度盐溶液的透析管道, 通过渗透脱水方式经一层聚二甲基硅氧烷(PDMS)膜实现液滴的逐渐浓缩, 使之趋于过饱和状态, 进而形成结晶. 此芯片可一次筛选较宽范围的过饱和状态, 实现蛋白质结晶的快速优化. 利用模式蛋白溶菌酶的结晶实验验证了该芯片的性能.
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| 关 键 词： | 自动进样 高通量筛选 渗透脱水 蛋白质结晶 |
| 收稿时间： | 2012-01-30 |
Microfluidic Device for Automated High-Throughput Protein Crystallization Screening by Osmotic Dewatering |
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| LUO Ya-Hui
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LI Gang
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CHEN Qiang
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ZHAO Jian-Long.Microfluidic Device for Automated High-Throughput Protein Crystallization Screening by Osmotic Dewatering[J].Chemical Research In Chinese Universities,2012,33(10):2178-2183. |
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| Authors: | LUO Ya-Hui LI Gang CHEN Qiang ZHAO Jian-Long |
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| Institution: | 1. State Key Lab of Transducer Technology, Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences, Shanghai 200050, China;
2. Graduate School of the Chinese Academy of Sciences, Beijing 100049, China |
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| Abstract: | A self-dispensing and osmosis-based microfluidic crystallization device was reported. This device automatically dispenses droplets containing protein and precipitant into an array of crystallization chambers by pre-degassing, and then gradually concentrates these droplets at different rates by osmotic dewatering through a polydimethylsiboxane(PDMS) membrane using a row of integrated dialysis channels filled with different concentration of salt solution. This concentration process drives the protein/precipitant mixture into supersaturated, thus tending toward the crystallization state. This device allows for screening a wide range of supersaturation in one trial for rapid optimization of protein crystallization. The feasibility of this crystallization device is demonstrated using the model proteins of lysozyme. |
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| Keywords: | Self-dispensing High-throughout screening Osmotic dewatering Protein crystallization |
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