کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
35903 | 45113 | 2007 | 6 صفحه PDF | دانلود رایگان |
The crystallization of lysozyme was monitored in 20-μl sitting-drop vapor diffusion experiments and a quantitative phase diagram was obtained. Then, batch crystallization of lysozyme in shaked 200-μl microtiter plates was investigated. It was observed that with rising agitation rates, the area of the nucleation zone was significantly reduced. Further batch crystallization experiments were performed (i) in 0.2–2-ml Eppendorf tubes in a laboratory rotator, (ii) in 5-ml unbaffled shake flasks, and (iii) in 4-ml stirred baffled and unbaffled vessels. The crystal area density distributions of the stirred vessels were clearly more narrow compared to the rotated Eppendorf tubes. The crystal area density distributions of the shake flasks were significantly wider. The use of the biocompatible, water-soluble ionic liquid ethanolammonium formate as a crystallization additive in unbaffled stirred vessels resulted in larger, sturdy crystals and reduced formation of crystal aggregates. The experiments indicate that ml-scale batch crystallization of lysozyme in stirred vessels can be performed fast, up-scaleable, reasonably reproducible, and precipitation can be avoided reliably.
Journal: Process Biochemistry - Volume 42, Issue 12, December 2007, Pages 1649–1654