Electric-Field-Assisted Protein Crystallization in Continuous Flow,Crystal Growth & Design

当前位置： X-MOL 学术 › Cryst. Growth Des. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Electric-Field-Assisted Protein Crystallization in Continuous Flow
Crystal Growth & Design ( IF 3.2 ) Pub Date : 2018-04-09 00:00:00 , DOI: 10.1021/acs.cgd.8b00095
Fei Li ₁ , Richard Lakerveld ₁

Affiliation

Crystallization is an attractive method for separation and purification of large biomolecules such as proteins due to inherent advantages related to product composition, stability, and process throughput. However, the lack of control over the nucleation and growth of protein crystals complicates design and operation. Electric fields have been used to improve control over the location and induction time of protein nucleation in small batches. To adopt electric-field-assisted protein crystallization as a separation and purification method, a novel device based on continuous flow is presented. The application of continuous flow allows for a large surface-to-volume ratio to be exploited for the electrode design and aligns with the current trend in the (bio)pharmaceutical industry toward continuous manufacturing. The device features millimeter-sized channels with coplanar electrodes to study the influence of nonuniform electric fields on protein crystallization in continuous flow. The results demonstrate that the induction time for protein nucleation can be reduced in continuous flow in comparison to batch crystallization irrespective of the application of an electric field. A higher yield can be attained for electric-field-assisted protein crystallization in continuous flow in comparison to control flow experiments without an electric field only when a longer residence time is applied, which is attributed to increased secondary nucleation at lower supersaturation. The device for electric-field-assisted protein crystallization in continuous flow can produce a large number of crystals of a small size, which is attractive for applications that require a large surface-to-volume ratio or for the use as seeds in downstream crystallizers optimized for crystal growth.

中文翻译：

连续流动中的电场辅助蛋白质结晶

由于与产物组成，稳定性和工艺产量有关的固有优势，结晶是用于分离和纯化诸如蛋白质的大生物分子的有吸引力的方法。但是，缺乏对蛋白质晶体成核和生长的控制，使设计和操作变得复杂。电场已用于改善对小批量蛋白质成核的位置和诱导时间的控制。以电场辅助蛋白结晶为分离纯化方法，提出了一种基于连续流的新型装置。连续流的应用允许将大的表面体积比用于电极设计，并与（生物）制药工业中朝着连续制造的当前趋势一致。该设备具有带有共面电极的毫米大小的通道，以研究不均匀电场对连续流动中蛋白质结晶的影响。结果表明，与分批结晶法相比，不管施加电场如何，在连续流动中都可以减少蛋白质成核的诱导时间。与不使用电场的控制流实验相比，仅当应用更长的停留时间时，与没有电场的控制流实验相比，连续流中的电场辅助蛋白质结晶才能获得更高的产量，这归因于较低的过饱和度下二次核的增加。电场辅助蛋白质在连续流中结晶的装置可以产生大量小尺寸的晶体，

更新日期：2018-04-09

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南