一种用于纯化在CHO细胞中产生的溶酶体贮积病蛋白β-葡萄糖醛酸酶的改进方法。
An improved purification method for the lysosomal storage disease protein β-glucuronidase produced in CHO cells.
作者信息
Fratz-Berilla Erica J, Ketcham Stephanie A, Parhiz Hamideh, Ashraf Muhammad, Madhavarao Chikkathur N
机构信息
Division of Product Quality Research, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Avenue, Silver Spring, MD 20993, USA.
Division of Product Quality Research, Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, Food and Drug Administration, 10903 New Hampshire Avenue, Silver Spring, MD 20993, USA.
出版信息
Protein Expr Purif. 2017 Dec;140:28-35. doi: 10.1016/j.pep.2017.07.011. Epub 2017 Jul 19.
Human β-glucuronidase (GUS; EC 3.2.1.31) is a lysosomal enzyme that catalyzes the hydrolysis of β-d-glucuronic acid residues from the non-reducing termini of glycosaminoglycans. Impairment in GUS function leads to the metabolic disorder mucopolysaccharidosis type VII, also known as Sly syndrome. We produced GUS from a CHO cell line grown in suspension in a 15 L perfused bioreactor and developed a three step purification procedure that yields ∼99% pure enzyme with a recovery of more than 40%. The method can be completed in two days and has the potential to be integrated into a continuous manufacturing scheme.
人β-葡萄糖醛酸酶（GUS；EC 3.2.1.31）是一种溶酶体酶，可催化从糖胺聚糖的非还原末端水解β-D-葡萄糖醛酸残基。GUS功能受损会导致代谢紊乱的VII型粘多糖贮积症，也称为斯利综合征。我们从在15升灌注生物反应器中悬浮培养的CHO细胞系中生产GUS，并开发了一种三步纯化程序，可产生纯度约为99%的酶，回收率超过40%。该方法可在两天内完成，并且有可能整合到连续生产方案中。
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