National Engineering Research Center of Industrial Crystallization Technology, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
National Engineering Research Center of Industrial Crystallization Technology, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China; Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300072, China.
Ultrason Sonochem. 2020 May;63:104975. doi: 10.1016/j.ultsonch.2020.104975. Epub 2020 Jan 18.
With the increasing demand for biopharmaceuticals, a method to crystallize biomolecule products with high quality, high yield and uniform size distribution as well as regular crystal habit is needed. In this work, ultrasound was used as a nucleation accelerator to decrease the energy barrier for lysozyme crystal formation. Crystallization experiments on egg-white lysozyme were carried out with and without ultrasound. The effect of ultrasound on induction time, metastable zone width, crystal size and morphology and process yield was investigated in detail. The nucleation-promoting effect produced by ultrasound is illustrated by the reduction of metastable zone width and induction time. By inducing faster nucleation, ultrasound leads to protein crystals grow at lower supersaturation levels with shorter induction time. It was found that ultrasound could result in uniform size distribution of the product due to the preventing of aggregation. However, long time continuous application of ultrasound could result in smaller particle size. Hence, ultrasonic-stop method was found to be a more appropriate strategy to enhance the crystallization process of proteins such as lysozyme.
随着对生物制药的需求不断增加，需要一种能够以高质量、高产率和均匀的尺寸分布以及规则的晶体习性来结晶生物分子产品的方法。在这项工作中，超声波被用作成核促进剂，以降低溶菌酶晶体形成的能量势垒。在有和没有超声波的情况下进行了蛋清溶菌酶的结晶实验。详细研究了超声波对诱导时间、亚稳区宽度、晶体尺寸和形态以及过程收率的影响。超声波产生的成核促进作用通过降低亚稳区宽度和诱导时间来体现。通过诱导更快的成核，超声波导致蛋白质晶体在较低的过饱和度水平下生长，具有更短的诱导时间。研究发现，由于防止聚集，超声波可以使产物的尺寸分布均匀。然而，长时间连续应用超声波会导致粒径变小。因此，发现超声停止法是一种更合适的策略，可以增强溶菌酶等蛋白质的结晶过程。
