CHO hücrelerinde monoklonal antikor üretim optimizasyonu ve karakterizasyonu

Biosimilar monoclonal antibody development is currently the major focus of the biopharmaceutical industry. Strategies to adjust the critical quality attributes early in the development stage saves time, labor, and costs considering the competitive global market. As a technologically and pharmaceutically developing country, particularly Turkey is currently expanding its biosimilars market and requires new developments. This study focuses on generating optimal conditions for the development of a biosimilar of C5mAb, in Chinese Hamster Ovary (CHO) cells. C5mAb acts as a blocker to the complement component 5, an important target for certain rare diseases. The novelties of the study are set forth with the usage of CHO host cell expression system to produce a hybrid and humanized mAb, the incorporation of Design of Experiments (DoEs) to screen and investigate process parameters and culture feeding regimens, and also by the process parameters optimized for the production of a C5mAb biosimilar. The study involves glycan and potency analyses as key quality attributes, with further verification in bioreactor systems, aiming to contribute to the assessment of biosimilarity. A robust 14 days fed-batch culture was developed with the E90-02 clone, under extended feed strategy. Applying temperature down-shift to 32°C from 37°C on the production day of 7, in stirred tank bioreactor system was confirmed to yield 2.9 g / L titer with 83.85 % binding potency, and matching glycosylation profiles, complying with the Originator molecule in terms of efficacy, safety, biological activity, immunogenicity, and stability. The study put forward the reliability and applicability of experimental design strategies for manufacturing bioprocesses that ensure compliance of key quality attributes, and contribute to the knowledge in the field of biosimilars development.