A microfluidic device for kinetic optimization of protein crystallization and in situ structure determination.
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.
Hansen
Carl L
CL
Department of Applied Physics, California Institute of Technology, 1200 East California Boulevard, Pasadena, California 91125, USA.
Classen
Scott
S
Berger
James M
JM
Quake
Stephen R
SR
D019747:Aldose-Ketose Isomerases / Q000737:chemistry; D002374:Catalase / Q000737:chemistry; D003460:Crystallization; D007700:Kinetics; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D015394:Molecular Structure; D009113:Muramidase / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D014961:X-Ray Diffraction
D010455:Peptides; D010940:Plant Proteins; D011506:Proteins; C019529:polyalanine; C011080:polyglycine; C003427:thaumatin protein, plant; D002374:Catalase; D009113:Muramidase; D019747:Aldose-Ketose Isomerases; C023298:xylose isomerase