Presentation on theme: "What We’ve Learned from the First 717,312 Crystallization Experiments George T. DeTitta 1, Melissa A. Bianca 1, Robert J. Collins 1, Ann Marie E. Faust."— Presentation transcript:
What We’ve Learned from the First 717,312 Crystallization Experiments George T. DeTitta 1, Melissa A. Bianca 1, Robert J. Collins 1, Ann Marie E. Faust 1, Jillian N. Kaczmarek 1, Joseph R. Luft 1, Nancy A. Urban 1, Walter A. Pangborn 1, Jennifer R. Wolfley 1, Igor Jurisica 2, Patrick Rogers 2, Gerald Quon 2, Janice Glasgow 3, Suzanne Fortier 3 (1) Hauptman-Woodward Medical Research Institute, 73 High Street, Buffalo, NY USA (2) Ontario Cancer Institute, Princess Margaret Hospital, University Health Network, Division of Cancer Informatics, 610 University Avenue, Room Toronto, Ontario M5G 2M9 Canada (3) Queen’s University, Kingston, Ontario K7L 3N6 Canada
Overview The HTP Crystallization Laboratory at HWI
Microbatch Under Oil Chayen, N.E., Stewart, P.D.S., Blow, D.M., Journal of Crystal Growth, 122 (1992)
The Experiment Plate Each plate contains one sample x 1536 unique chemical cocktails
Setting Up the Plates Source/Destination Plate Methods Reformat Plates from Low to High Density Modified Robbins Scientific Hydra 96 and 384 are used to deliver the solutions
Hydra 384 Liquid Handling System
Imaging the Plates
Enlarged Image with Details
Animated Views 1536 different Bovine liver catalase crystallization experiments were setup. Every experiment was recorded 203 times at 30 minute intervals. Still images were used to produce time- lapse recordings of the experiments. Determine preliminary outcomes that could lead to crystallization.
Underlying Physical Parameters Activity of water Viscosity Surface Tension pH Ionic Strength Hofmeister Series Excluded Volume Specific Ion Effects
Ancillary Data Collection Techniques used to characterize proteins SDS-PAGE Dynamic Light Scattering Instruments used to characterize cocktails Refractometer Vapor Pressure Osmometer Tensiometer Viscometer
Examples of Outcomes
User Profile GroupLaboratory Use Structural biology54.8% Structural genomics38.5% Pharmaceutical 6.7%
Conclusions HTP crystallization can have dual roles: 1) A method to produce crystals 2) A tool for knowledge discovery Setting up a lot of crystallization experiments is not a cure-all (GIGO). Method variation can produce different results. Image quality is critical for success. Verification of the outcomes’ composition is an important focus for development.
Acknowledgements John R. Oishei Foundation Cummings Foundation NASA NAG NIH grant 1 P50 GM NIH grant 1 P50 GM IBM SUR NSERC (Natural Sciences and Engineering Research Council of Canada) CITO (Communications and Information Technology Ontario)