间歇式结晶器中采用过程分析技术的蛋白质成核和结晶过程,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.)

间歇式结晶器中采用过程分析技术的蛋白质成核和结晶过程
Crystal Growth & Design ( IF 3.2 ) Pub Date : 2023-06-20 , DOI: 10.1021/acs.cgd.3c00411
Wenqing Tian ₁ , Wei Li ₁ , Huaiyu Yang ₁

Affiliation

蛋白质结晶因其在稳定性、储存和递送方面的优势而受到广泛关注，以替代蛋白质药物的传统下游加工。对蛋白质结晶过程的了解有限，需要基于结晶过程中实时跟踪的基本信息。配备聚焦光束反射测量 (FBRM) 探头和热电偶的 100 mL 批量结晶器设计用于原位监测蛋白质结晶过程，同时记录离线浓度和晶体图像。确定了蛋白质批量结晶过程的三个阶段：长周期缓慢成核、快速结晶以及缓慢生长和破裂。通过FBRM估计诱导时间，即溶液中颗粒数量的增加，这可能是通过离线测量检测浓度下降所需时间的一半。在相同盐浓度下，诱导时间随着过饱和度的增加而缩短。分析了等盐浓度和不同溶菌酶浓度的各实验组的成核界面能。界面能随着溶液中盐浓度的增加而降低。实验的产率受到蛋白质和盐浓度的显着影响，在浓度读数稳定后，产率高达 99%，中值晶体尺寸为 26.5 μm。在相同盐浓度下，诱导时间随着过饱和度的增加而缩短。分析了等盐浓度和不同溶菌酶浓度的各实验组的成核界面能。界面能随着溶液中盐浓度的增加而降低。实验的产率受到蛋白质和盐浓度的显着影响，在浓度读数稳定后，产率高达 99%，中值晶体尺寸为 26.5 μm。在相同盐浓度下，诱导时间随着过饱和度的增加而缩短。分析了等盐浓度和不同溶菌酶浓度的各实验组的成核界面能。界面能随着溶液中盐浓度的增加而降低。实验的产率受到蛋白质和盐浓度的显着影响，在浓度读数稳定后，产率高达 99%，中值晶体尺寸为 26.5 μm。界面能随着溶液中盐浓度的增加而降低。实验的产率受到蛋白质和盐浓度的显着影响，在浓度读数稳定后，产率高达 99%，中值晶体尺寸为 26.5 μm。界面能随着溶液中盐浓度的增加而降低。实验的产率受到蛋白质和盐浓度的显着影响，在浓度读数稳定后，产率高达 99%，中值晶体尺寸为 26.5 μm。

"点击查看英文标题和摘要"

Protein Nucleation and Crystallization Process with Process Analytical Technologies in a Batch Crystallizer

Protein crystallization has drawn great attention to replacing the traditional downstream processing for protein-based pharmaceuticals due to its advantages in stability, storage, and delivery. Limited understanding of the protein crystallization processes requires essential information based on real-time tracking during the crystallization process. A batch crystallizer of 100 mL fitted with a focused beam reflectance measurement (FBRM) probe and a thermocouple was designed for in situ monitoring of the protein crystallization process, with simutaneously record of off-line concentrations and crystal images. Three stages in the protein batch crystallization process were identified: long-period slow nucleation, rapid crystallization, and slow growth and breakage. The induction time was estimated by FBRM, i.e., increasing numbers of particles in the solution, which could be half of the time required for detecting the decrease of the concentration, by offline measurement. The induction time decreased with an increase in supersaturation within the same salt concentration. The interfacial energy for nucleation was analyzed based on each experimental group with equal salt concentration and different concentrations of lysozyme. The interfacial energy reduced with an increase in salt concentration in the solution. The yield of the experiments was significantly affected by the protein and salt concentrations and could achieve up to 99% yield with a 26.5 μm median crystal size upon stabilized concentration readings.

更新日期：2023-06-20

点击分享查看原文

点击收藏

公开下载

阅读更多本刊新发论文本刊介绍/投稿指南