Design, 3D printing and validation of a novel low‐cost high‐capacity sitting‐drop bridge for protein crystallization

• Published in: Journal of applied crystallography 2019-02, Vol.52 (1), p.171-174
• Main Authors: Talapatra, Sandeep K., Penny, Matthew R., Hilton, Stephen T., Kozielski, Frank
• Format: Article
• Language: English
• Subjects: 3-D printers; 3D printing; Crystallization; hanging‐drop crystallization; microbridges; Optimization; protein crystallization; Proteins; sitting‐drop crystallization; Three dimensional printing; vapour‐diffusion crystallization
• ISSN: 1600-5767; 0021-8898; 1600-5767
• DOI: 10.1107/S1600576718017545

Sitting‐drop protein crystallization is not used as commonly as the hanging‐drop method for crystal optimization owing to the limitations of commercially available sitting‐drop bridges, particularly when they are used in conjunction with 24‐well crystallization plates. The commercially available sitting‐drop bridge, containing space for only a single drop, restricts their wider use. Proteins that preferentially crystallize under sitting‐drop conditions therefore require more work, time and resources for their optimization. In response to these limitations, and using 3D printing, a new sitting‐drop bridge has been designed and developed, where five crystallization drops can be placed simultaneously in each well of a 24‐well crystallization plate. This significantly simplifies the process and increases the potential of sitting drops in crystal optimization, reducing costs and hence overcoming the limitations of current approaches. Optimization of protein crystals using the vapour‐diffusion sitting‐drop method is time consuming, labour intensive and costly. This article presents the concept, design and evaluation of a novel low‐cost and high‐capacity sitting‐drop bridge. It is hoped that this will help to remove the present bottleneck in crystal optimization and streamline the overall process to obtain high‐quality crystals.