229f High Throughput Nanoliter Screening and Morphological Control of Protein Crystals Via Precipitant Gradients

Goher Mahmud, Stoyan K. Smoukov, Yuriy Chegel, and Bartosz A. Grzybowski. Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Ave., Tech E-136, Evanston, IL 60208

With the high cost of proteins, and the numerous experiments that must be run in order to determine correct crystallization conditions, it is imperative that the screening process use small quantities to be effective. Here, we present a method that uses nanoliter volumes of protein that can develop temporal and spatial precipitant gradients, which allows for screening of many conditions with resolution not even considered for model protein crystallization. Also, we show that the large number of small wells allows for measurement of statistical quantities of nucleation and control of crystal polymorphism. Finally, thermodynamic models of crystal face growth were combined with measurements of crystal nucleation rates, protein super saturation levels, and crystal face growth rates to optimize crystallization of particular polymorphs.


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