Three-dimensional structure determination using X-ray crystallography is a linear process, and failure to complete any step successfully will terminate a structural study. Crystal growth has long been identified as a bottleneck. There are currently no rational means for predicting good crystallization conditions for a particular macromolecule. Many experiments are set up in the hopes that one will lead to suitable results. The methods employed are often tedious and inefficient, and they fail to thoroughly explore all of the possible chemical and physical conditions.

At HWI, a group led by Dr. George DeTitta and Mr. Joseph Luft has developed an automated method to set up high-throughput crystallization experiments. As of March 2011, more than 21 million experiments have been performed, and the results recorded in the form of ~120 million digital images. More than 1000 researchers have provided samples of 14,000 proteins and nucleic acids for these experiments. The supplier receives a set of digital images showing the outcomes of 1536 unique crystallization trials. The data generated from these experiments is being used to create a database that we hope will permit better crystallization conditions for new macromolecules to be predicted rationally. The high-throughput lab is developing tools for crystallographic structure determination from sample preparation to cryogenically preserved, mounted crystals ready for X-ray diffraction experiments. Recent publications include work by Dr. Eddie Snell to chemically map and predict how to optimize crystallization conditions based upon results from the 1536 screening experiments.

It is often necessary to fine-tune initial crystallization conditions in order to obtain the high quality crystals required for structure determination. Studies at HWI have led to the development of techniques to further this process. One method, seed bead, is commercially available.  A diffusion plate for growing crystals and a method for high-throughput crystallization screening have been separately patented.

We gratefully acknowledge the past support of: The Cummings Foundation, The John R. Oishei Foundation, The Western New York Foundation, NASA NAG8-1594, NASA NAG8-1839, NASA NCC8-232, NIH GM-64655, NIH GM-62413 and NIH NCRR S10 RR016924.

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