Efficient optimization of crystallization conditions by manipulation of drop volume ratio and temperature

• Published in: Protein science Vol. 16; no. 4; pp. 715 - 722
• Main 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.
• Format: Journal Article
• Language: English
• Published: Bristol Cold Spring Harbor Laboratory Press 01-04-2007
• Subjects: Crystallization; high throughput; optimization; protein crystallization; Robotics; Temperature
• ISSN: 0961-8368; 1469-896X
• DOI: 10.1110/ps.062699707

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.