Combining Counter-Diffusion and Microseeding to Increase the Success Rate in Protein Crystallization

• Published in: Crystal growth & design 2011-06, Vol.11 (6), p.2122-2126
• Main Authors: Gavira, Jose A, Hernandez-Hernandez, M. A, Gonzalez-Ramirez, Luis A, Briggs, Richard A, Kolek, Stefan A, Shaw Stewart, Patrick D
• Format: Article
• Language: English
• Subjects: Condensed matter: structure, mechanical and thermal properties; Cross-disciplinary physics: materials science; rheology; Diffusion in solids; Equations of state, phase equilibria, and phase transitions; Exact sciences and technology; General studies of phase transitions; Materials science; Methods of crystal growth; physics of crystal growth; Nucleation; Physics; Solid-solid transitions; Specific phase transitions; Transport properties of condensed matter (nonelectronic)
• ISSN: 1528-7483; 1528-7505
• DOI: 10.1021/cg101380j

A new method of increasing the success rate in protein crystallization screening experiments by combining microseeding with counter-diffusion crystallization in capillaries (SCD) is presented. We have investigated the number of crystallization hits obtained with and without microseeding with 10 model proteins. For the cases studied, SCD generally increases the number of hits and is particularly useful when only relatively low protein concentration stocks are available, either because the stocks were prepared for, e.g., vapor diffusion experiments, or because the protein is poorly soluble. In either case, the addition of seeds becomes necessary to overcome the nucleation energy barrier so that crystal growth can take place even when the wave of supersaturation that passes along the capillary is insufficient to promote nucleation.