Automated High-Throughput Protein Crystallization
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X-ray crystallography provides key biological insights into the three-dimensional structure and function of proteins, as well as essential information on protein-protein and protein-ligand interactions, therefore facilitating the design of more effective clinical drugs. The three most popular protein crystallization methods—vapor-diffusion sitting drop, hanging drop, and microbatch (1, 2, 3, 4, 5, 6), most commonly used by investigators-convey major economical disadvantages for the setup of rapid large-scale crystallization of new proteins. Lack of suitable automation for the numerous lengthy and labor-intensive setup steps, irreproducibility as a result of manual intervention, waste of precious and scarce protein samples caused by the absence of precise low-volume dispensers and appropriate plate technology, exorbitant consumption of time, and cost (4,5) are among the most common drawbacks of high-throughput crystallization.
KeywordsProtein Crystallization Crystallization Reaction Plate Technology Screen Plate Viscous Protein
- 1.McPherson, A. (1982) Preparation and analysis of protein crystals. Krieger, Malabar, FL.Google Scholar
- 2.Rhodes, G. (2000) Crystallography made crystal clear: a guide for users of macromolecular models. Academic, San Diego, CA.Google Scholar
- 3.Hansen, L., Skordalakes, E., Berger, J. M., and Quake, S. R. (2002) A robust and scalable microfluidic metering method that allows protein crystal growth by free interface diffusion. Proc. Natl. Acad. Sci. USA 99, 16,531-16,536.Google Scholar
- 7.Krupka, H. I., Rupp, Segelke, B. W., et al. (2002) The high-speed Hydra®-Plus-One system for automated high-throughput protein crystallography. Acta. Cry st. D53,1523–1526.Google Scholar
- 8.James, A., Wu, H.-C., Braunthal, N., Shieh, J., and Azarani, A. (2003) Setting Up high-throughput, low-volume sequencing and PCR reactions using an automated system equipped with precision glass syringes and a non-contact microsolenoid dispenser. JALA 8, 37–40.Google Scholar
- 9.Stanchfield, J., Wright, D., Hsu, S., Lamsa, M., and Robbins, A. (1996) Precision 96-channel dispenser for microchemical techniques. BioTechniques 20, 292–296.Google Scholar
- 10.Mardis, E. R., Weinstock, L., Simonyan, A., and Stanchfield, J. E. (1997) M13 DNA prepa-rations for a large scale sequencing project using the Hydra-96 microdispenser. Product Application Note 5, Apogent Discoveries (Web site at http://www.apogentdiscoveries.com).
- 12.To, C., Todd, P., Wright, D., and Azarani, A. (2001) Prevention of carry-over contamination from organic compounds, DNA and protein samples when using the Robbins Tango Liquid Handling System. Product Application Note 10, Apogent Discoveries (web site at http://www.apogentdiscoveries.com).
- 14.Azarani, A. (2004) The use of precision glass syringes and a noncontact microsolenoid dispenser for the production of high-throughput low-density arrays. In: (eiFung, E. T., ed.) Protein Arrays: Methods and Protocols. Humana, Totowa, NJ.Google Scholar
- 15.James, A. T., Wu, H.-C., Braunthal, N., and Azarani, A. (2003) A study of a high-through-put plasmid DNA purification system, JALA 8, 36–39.Google Scholar