Lessons from high-throughput protein crystallization screening: 10 years of practical experience

Abstract

Introduction: X-ray crystallography provides the majority of our structural biological knowledge at a molecular level and, in terms of pharmaceutical design, is a valuable tool to accelerate discovery. It is the premier technique in the field, but its usefulness is significantly limited by the need to grow well-diffracting crystals. It is for this reason that high-throughput crystallization has become a key technology that has matured over the past 10 years through the field of structural genomics.

Areas covered : The authors describe their experiences in high-throughput crystallization screening in the context of structural genomics and the general biomedical community. They focus on the lessons learnt from the operation of a high-throughput crystallization-screening laboratory, which to date has screened over 12,500 biological macromolecules. They also describe the approaches taken to maximize the success while minimizing the effort. Through this, the authors hope that the reader will gain an insight into the efficient design of a laboratory and protocols to accomplish high-throughput crystallization on a single-, multiuser laboratory or industrial scale.

Expert opinion : High-throughput crystallization screening is readily available but, despite the power of the crystallographic technique, getting crystals is still not a solved problem. High-throughput approaches can help when used skillfully; however, they still require human input in the detailed analysis and interpretation of results to be more successful.

Keywords::

• crystallization
• high-throughput
• structural biology
• X-ray crystallography

Acknowledgments

The members of the crystallization-screening laboratory are acknowledged for their tireless efforts to maintain an efficient laboratory. We thank B Cudney for his informative discussions and insights. The many users of the screening laboratory are also thanked, in particular G Montelione from the NESG for access to detailed information about all of the samples run through the screening laboratory.

Notes

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