W0470

HTS for Crystallization Conditions Achilles Heel: Positive Identification of Lead Conditions. Joseph Luft^1,2, Robert Collins¹, Nancy Fehrman¹, Angela Lauricella¹, Christina Veatch¹, George DeTitta^1,2, ¹Hauptman-Woodward Medical Research Institute; ²Dept. of Structural Biology, SUNY at Buffalo.

A facility was established for high throughput screening of initial crystallization conditions at the Hauptman-Woodward Institute in February 2000. The facility uses the microbatch-under-oil method (N. E. Chayen, P. D. Shaw Stewart, and D. M. Blow. (1992) J. Crystal Growth 122, 176-180.) to set up 1536 well experiment plates. Each plate contains one macromolecular sample combined with 1536 unique crystallization solutions. The outcomes of the experiments are recorded for a period of one month. Images are available to users of the facility through a secure ftp server. As of May 2003, the laboratory was used to set up 1868 samples (2.9 million crystallization trials; 19 million images) for 350 researchers. The ability to set up and record millions of experiments designed to identify initial crystallization conditions creates a new bottleneck: verification of lead conditions. Although it is possible to recognize “crystals” from image data, it is not possible to identify their composition (salt or macromolecule). Experiments are underway to identify compounds that will enable verified identification of macromolecular crystallization lead conditions. The evolution of technology in the HTS laboratory along with statistics on the experiments will be presented. Work was supported in part by NIH GM-64655 (SGPP), GM-62413 (NESG), NCRR S10 RR016924, NASA NAG8-1594, NAG8-1839, NCC8-232, The John R. Oishei Foundation, The Cummings Foundation, and The Western New York Foundation.