Efficient optimization of crystallization conditions by manipulation of drop volume ratio and temperature |
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Authors: | Luft Joseph R Wolfley Jennifer R Said Meriem I Nagel Raymond M Lauricella Angela M Smith Jennifer L Thayer Max H Veatch Christina K Snell Edward H Malkowski Michael G Detitta George T |
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Affiliation: | Hauptman-Woodward Medical Research Institute, Buffalo, New York 14203, USA. luft@hwi.buffalo.edu |
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Abstract: | An efficient optimization method for the crystallization of biological macromolecules has been developed and tested. This builds on a successful high-throughput technique for the determination of initial crystallization conditions. The optimization method takes an initial condition identified through screening and then varies the concentration of the macromolecule, precipitant, and the growth temperature in a systematic manner. The amount of sample and number of steps is minimized and no biochemical reformulation is required. In the current application a robotic liquid handling system enables high-throughput use, but the technique can easily be adapted in a nonautomated setting. This method has been applied successfully for the rapid optimization of crystallization conditions in nine representative cases. |
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Keywords: | protein crystallization high throughput optimization temperature |
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