A procedure for setting up high-throughput nanolitre crystallization experiments. II. Crystallization results

• Published in: Journal of applied crystallography Vol. 36; no. 2; pp. 315 - 318
• Main Authors: Brown, J., Walter, T. S., Carter, L., Abrescia, N. G. A., Aricescu, A. R., Batuwangala, T. D., Bird, L. E., Brown, N., Chamberlain, P. P., Davis, S. J., Dubinina, E., Endicott, J., Fennelly, J. A., Gilbert, R. J. C., Harkiolaki, M., Hon, W.-C., Kimberley, F., Love, C. A., Mancini, E. J., Manso-Sancho, R., Nichols, C. E., Robinson, R. A., Sutton, G. C., Schueller, N., Sleeman, M. C., Stewart-Jones, G. B., Vuong, M., Welburn, J., Zhang, Z., Stammers, D. K., Owens, R. J., Jones, E. Y., Harlos, K., Stuart, D. I.
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England Munksgaard International Publishers 01.04.2003; Blackwell
• Subjects: Biological and medical sciences; Cross-disciplinary physics: materials science; rheology; crystallization; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; Growth from solutions; In solution. Condensed state. Thin layers; ink-jet technology; macromolecular crystallography; Materials science; Methods of crystal growth; physics of crystal growth; Molecular biophysics; Physico-chemical properties of biomolecules; Physics; protein crystallography; Crystallization; Biological macromolecule; XRD; Evaporation; Crystal perfection; Robotics; Proteins; Nanometer scale; Monocrystals; Sample preparation; Automatic system; Crystal growth from solutions; Droplets; Sitting drop method; Synchrotron radiation; Crystal structure
• ISSN: 1600-5767; 0021-8898; 1600-5767
• DOI: 10.1107/S0021889803002012

An initial tranche of results from day‐to‐day use of a robotic system for setting up 100 nl‐scale vapour‐diffusion sitting‐drop protein crystallizations has been surveyed. The database of over 50 unrelated samples represents a snapshot of projects currently at the stage of crystallization trials in Oxford research groups and as such encompasses a broad range of proteins. The results indicate that the nanolitre‐scale methodology consistently identifies more crystallization conditions than traditional hand‐pipetting‐style methods; however, in a number of cases successful scale‐up is then problematic. Crystals grown in the initial 100 nl‐scale drops have in the majority of cases allowed useful characterization of X‐ray diffraction, either in‐house or at synchrotron beamlines. For a significant number of projects, full X‐ray diffraction data sets have been collected to 3 Å resolution or better (either in‐house or at the synchrotron) from crystals grown at the 100 nl scale. To date, five structures have been determined by molecular replacement directly from such data and a further three from scale‐up of conditions established at the nanolitre scale.