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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.

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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:

1 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

2 Overview The HTP Crystallization Laboratory at HWI

3 Microbatch Under Oil Chayen, N.E., Stewart, P.D.S., Blow, D.M., Journal of Crystal Growth, 122 (1992)

4 The Experiment Plate Each plate contains one sample x 1536 unique chemical cocktails

5 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

6 Hydra 384 Liquid Handling System

7 Imaging the Plates

8 MacroScope

9 Enlarged Image with Details

10 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.

11 Improvements  Decreased Solution Volume  Cocktails  Image Quality  Image Viewing and Sorting  Image Analysis

12 Underlying Physical Parameters  Activity of water  Viscosity  Surface Tension  pH  Ionic Strength  Hofmeister Series  Excluded Volume  Specific Ion Effects

13 Ancillary Data Collection Techniques used to characterize proteins  SDS-PAGE  Dynamic Light Scattering Instruments used to characterize cocktails  Refractometer  Vapor Pressure Osmometer  Tensiometer  Viscometer

14 Examples of Outcomes

15 User Profile GroupLaboratory Use Structural biology54.8% Structural genomics38.5% Pharmaceutical 6.7%

16 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.

17 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)


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