The critical role of agitation in moving from preliminary screening results to reproducible batch protein crystallisation
The critical role of agitation in moving from preliminary screening results to reproducible batch protein crystallisation
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DOI：
10.1016/j.cherd.2021.06.012
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发表时间：
2021-07-16
影响因子：
3.9
通讯作者：
Heng, Jerry Y. Y.
中科院分区：
文献类型：
--
作者：
Li, Xiaoyu;Heng, Jerry Y. Y.
本研究探讨了从蛋白质结晶的初步定性筛选到定量分批结晶实验时，搅拌在获得一致和可重现结果方面的重要作用。本研究以溶菌酶-索马甜二元蛋白混合物为模型蛋白体系。即使采用相同的采样时间和频率，在非搅拌结晶条件下也观察到批次间重现性较差。在本研究中研究的0至200 rpm搅拌条件下，随着搅拌的增加，观察到蛋白质结晶的重现性改善。此外，搅拌对过饱和度消耗速率、收率和晶体尺寸也有影响。此外，在搅拌分批结晶中，发现在蛋白质杂质的存在下，目标蛋白质结晶过程减慢。总之，我们强调了搅拌在蛋白质结晶实验中的重要作用，否则可能会从非搅拌系统中得出不一致的误导性结论。(c)2021化学工程师学会。Elsevier B. V.出版，保留所有权利。
This study investigated the important role of agitation in obtaining consistent and reproducible results when moving from preliminary qualitative screenings for protein crystallisation to quantitative batch crystallisation experiments. Lysozyme-thaumatin binary protein mixture was used as the model protein system in this study. Poor reproducibility between batches were observed for non-agitated crystallisation conditions even if the same sampling timing and frequency applied. With agitation, from 0 to 200 rpm investigated in this study, improved reproducibility of protein crystallisation was observed with increased agitation. Additionally, agitation also had impacts on supersaturation exhaustion rate, yield and crystal size. Moreover, in agitated batch crystallisation, it was found that target protein crystallisation process was decelerated in the presence of protein impurity. In conclusion, we emphasised the essential role of agitation in protein crystallisation experiments else misleading conclusions with inconsistency might be drawn from non-agitated systems. (c) 2021 Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.