Abstract: Provided is a method for large-scale production of lentivirus by using GMP-level serum-free suspension cells. Said method comprises the following steps: (a) providing a seed solution of packaged cells; (b) inoculating the seed solution in a first culture solution; (c) carrying out subculture of the packaged cells; (d) starting a liquid change operation when a liquid change trigger condition is met; (e) repeating steps (c) and (d) 1, 2 or 3 times; (f) starting a transfection operation when a transfection trigger condition is met; (g) optionally performing liquid change after transfection; (h) cultivating the transfected packaged cells; (i) starting harvesting and liquid change operations when a liquid change trigger condition is met; (j) repeating steps (h) and (i) 1, 2 or 3 times; (k) combining each of the recovered liquids; and (l) performing a purifying treatment. The culture solution used in each step is a serum-free cell culture solution.
Type:
Application
Filed:
October 31, 2023
Publication date:
February 22, 2024
Applicant:
Shanghai Cellular Biopharmaceutical Group Ltd.
Inventors:
Yi HONG, Ting YAN, Jiangguo YING, Haojie ZHANG, Luyi ZHANG, Li ZHANG
Abstract: Provided is a method for large-scale production of lentivirus by using GMP-level serum-free suspension cells. Said method comprises the following steps: (a) providing a seed solution of packaged cells; (b) inoculating the seed solution in a first culture solution; (c) carrying out subculture of the packaged cells; (d) starting a liquid change operation when a liquid change trigger condition is met; (e) repeating steps (c) and (d) 1, 2 or 3 times; (f) starting a transfection operation when a transfection trigger condition is met; (g) optionally performing liquid change after transfection; (h) cultivating the transfected packaged cells; (i) starting harvesting and liquid change operations when a liquid change trigger condition is met; (j) repeating steps (h) and (i) 1, 2 or 3 times; (k) combining each of the recovered liquids; and (1) performing a purifying treatment. The culture solution used in each step is a serum-free cell culture solution.
Type:
Grant
Filed:
March 28, 2019
Date of Patent:
December 19, 2023
Assignee:
SHANGHAI CELLULAR BIOPHARMACEUTICAL GROUP LTD.
Inventors:
Yi Hong, Ting Yan, Jiangguo Ying, Haojie Zhang, Luyi Zhang, Li Zhang
Abstract: Provided is a method for large-scale preparation of a purified preparation of a recombinant lentiviral vector at the GMP grade. The method comprises: (a) providing raw material feed liquid to be purified that comprises recombinant viral vectors; (b) carrying out a microfiltration treatment on the feed liquid to obtain a microfiltered filtrate comprising the recombinant viral vectors; (c) optionally concentrating the filtrate to obtain a concentrated filtrate; (d) purifying the filtrate obtained in the previous step by means of chromatography to obtain a crude pure product comprising the recombinant viral vectors; and (e) subjecting the crude pure product obtained in the previous step to liquid exchange and elaborate purification to obtain the purified recombinant viral vectors.
Type:
Application
Filed:
March 28, 2019
Publication date:
January 14, 2021
Applicants:
SHANGHAI CELLULAR BIOPHARMACEUTICAL GROUP LTD., WUXI CELLULAR BIOPHARMACEUTICAL GROUP LTD.
Inventors:
Yi Hong, Ting Yan, Jiangguo Ying, Haojie Zhang, Li Zhang, Fei Wang, Dijun Zhao, Luyi Zhang