Optimized Upstream Process Development for Production of a Leunase (L-Asparaginase) Biosimilar
Volume Title: 1
1Sasan Jahanshahi; 2Arezou Najafikhah; 3Leila Barani; 3Farzad Khanipour; 4Zohre Fathinezhad; 5Morteza Jafaraghaei; 6Amir Maghsoudi
1Microbial cell culture Laboratory, R&D Department, PersisGen Par Company.
2Microbial cell culture Laboratory, Production Department, PersisGen Par Company.
3Scientific and technical information, R&D Depatment, PersisGen Par Company
4Physicochemical Laboratory, Quality Control Department, PersisGen Par Company
5Deputy Director, R&D Department, PersisGen Par Company
6Manager, R&D Department, PersisGen Par Company
Asparaginase is an enzyme that catalyzes the hydrolysis of L-Asparagine to L-Aspartic acid and ammonia which is one of the most significant oncologic drugs for the treatment of leukemia. Therefore, process developments for pharmaceutical-grade production of L-Asparaginase biosimilars is being pursued worldwide. In this research, cell culture process for production of an L-Asparaginase biosimilar candidate, amidohydrolase type II expressed in E.coli host, was developed regarding the quality-by-design (QbD) principles. Also design of experiments, based on risk assessment of the production process, was carried out to control the critical process parameters (CPPs) which could significantly affect the final product quality. Based on the results of cell culture process optimization, feeding of glucose %50, µset = 0.13 h-1 and IPTG as the inducer for expression of the protein product, were selected. SDS-PAGE results showed an appropriate amount of protein expression after cell culture induction with IPTG. Also, the enzyme activity assays demonstrated that enzymatic activity of the biosimilar product sample was within acceptable range with ~106% of activity relative to that of the reference standard drug samples.