期刊
SENSORS AND ACTUATORS B-CHEMICAL
卷 174, 期 -, 页码 1-9出版社
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2012.08.048
关键词
X-ray transparency; Microfabrication; Protein crystallization; Structure determination; Crystallography
资金
- NIH [R01 GM086727]
- NIH from National Institute of Biomedical Imaging and Bioengineering [F31 EB008330]
- U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences [DE-AC02-06CH11357]
- Michigan Economic Development Corporation
- Michigan Technology Tri-Corridor [085P1000817]
This paper reports a method for fabricating multilayer microfluidic protein crystallization platforms using different materials to achieve X-ray transparency and compatibility with crystallization reagents. To validate this approach, three soluble proteins, lysozyme, thaumatin, and ribonuclease A were crystallized on-chip, followed by on-chip diffraction data collection. We also report a chip with an array of wells for screening different conditions that consume a minimal amount of protein solution as compared to traditional screening methods. A large number of high quality isomorphous protein crystals can be grown in the wells, after which slices of X-ray data can be collected from many crystals still residing within the wells. Complete protein structures can be obtained by merging these slices of data followed by further processing with crystallography software. This approach of using an X-ray transparent chip for screening, crystal growth, and X-ray data collection enables room temperature data collection from many crystals mounted in parallel, which thus eliminates crystal handling and minimizes radiation damage to the crystals. (C) 2012 Elsevier B.V. All rights reserved.
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