What We’ve Learned from the First 717,312 Crystallization Experiments George T. DeTitta1, Melissa A. Bianca1, Robert J. Collins1, Ann Marie E. Faust1, Jillian N. Kaczmarek1, Joseph R. Luft1, Nancy A. Urban1, Walter A. Pangborn1, Jennifer R. Wolfley1, Igor Jurisica2, Patrick Rogers2, Gerald Quon2, Janice Glasgow3, Suzanne Fortier3

  (1)   Hauptman-Woodward Medical Research Institute, 73 High Street, Buffalo, NY 14203-1196 USA (2)   Ontario Cancer Institute, Princess Margaret Hospital, University Health Network, Division of Cancer Informatics, 610 University Avenue, Room 8-413 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 CrystalGrowth, 122 (1992) 176-180.

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

MacroScope

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.

Improvements

Decreased Solution VolumeCocktailsImage QualityImage Viewing and SortingImage Analysis

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

Ancillary Data CollectionTechniques used to characterize proteins SDS-PAGE Dynamic Light Scattering

Instruments used to characterize cocktails Refractometer Vapor Pressure Osmometer Tensiometer Viscometer

Examples of Outcomes

User ProfileGroup Laboratory UseStructural biology 54.8%Structural genomics

38.5%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 NAG8-1594 NIH grant 1 P50 GM62413-01 NIH grant 1 P50 GM64655-01 IBM SUR NSERC (Natural Sciences and Engineering

Research Council of Canada) CITO (Communications and Information

Technology Ontario)