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Communication

A Microfluidic Device for Kinetic Optimization of Protein Crystallization and In Situ Structure Determination

Supporting Info

Carl L. Hansen,^† Scott Classen,^† James M. Berger,^† and Stephen R. Quake*^†

Department of Applied Physics, California Institute of Technology, 1200 East California Boulevard, Pasadena, California 91125

J. Am. Chem. Soc., 2006, 128 (10), pp 3142–3143

DOI: 10.1021/ja0576637

Publication Date (Web): February 15, 2006

†

Current addresses: (C.L.H.) Physics & Astronomy and Electrical & Computer Engineering, The University of British Columbia, Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T-1Z4 Canada. (S.C.) Advanced Light Source, Lawrence Berkeley National Laboratory, 1 Cyclotron Road Mail Stop 6R2100, Berkeley, CA 94720-8226, USA. (J.M.B.) Molecular and Cell Biology, 327B Hildebrand Hall #3206, University of California at Berkeley, Berkeley, CA 94720-3206, USA. (S.R.Q.) Bioengineering, Stanford University, Stanford, CA 94305, USA.

In papers with more than one author, the asterisk indicates the name of the author to whom inquiries about the paper should be addressed.

, quake@stanford.edu

Abstract

The unprecedented economies of scale and unique mass transport properties of microfluidic devices made them viable nano-volume protein crystallization screening platforms. However, realizing the full potential of microfluidic crystallization requires complementary technologies for crystal optimization and harvesting. In this paper, we report a microfluidic device which provides a link between chip-based nanoliter volume crystallization screening and structure analysis through “kinetic optimization” of crystallization reactions and in situ structure determination. Kinetic optimization through systematic variation of reactor geometry and actuation of micromechanical valves is used to screen a large ensemble of kinetic trajectories that are not practical with conventional techniques. Using this device, we demonstrate control over crystal quality, reliable scale-up from nanoliter volume reactions, facile harvesting and cryoprotectant screening, and protein structure determination at atomic resolution from data collected in-chip.

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