Method for regulating protein charge heterogeneity by ATF perfusion culture process

• Main Authors: YANG HUIMIN, LI ZHI, SHAN LIYUAN, ZHU GEXIN, CHENG JIN, WU QIWEI, XU XIAOFANG
• Format: Patent
• Language: Chinese; English
• Published: 08.08.2023
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

The invention discloses a method for adjusting protein charge heterogeneity through an ATF perfusion culture process, and belongs to the technical field of biology. According to the method, cells and expressed target products are intercepted in a reactor in a perfusion culture mode, metabolic waste is discharged through a 50KD ATF column, and the purpose of increasing the protein yield is achieved while charge acidic components in a culture environment are reduced. Compared with a traditional method for adjusting charge heterogeneity, the method has the advantages that the protein yield is increased by two times, the acidic component of the charge heterogeneity is reduced from 32.51% to 21.5-22.7%, and the alkaline component is increased from 11.03% to 21.7-22.9%. 本发明公开了一种ATF灌流培养工艺调节蛋白电荷异质性的方法，属于生物技术领域。本发明通过灌流培养的方式将细胞和表达的目的产物截留在反应器中，代谢废物通过50KD的ATF柱排出，在降低培养环境中的电荷酸性成分的同时，达到提高蛋白产量的目的。相较于传统调节电荷异质性的方法，蛋白产量提高了2倍，电荷异质性的酸性组分从32.51%降低至21.5~22.7%，碱性组分从11.03%提高至21.7~22.9%。