GMP Phase 1 Clinical Production of Therapeutic Antibodies
Upstream Process – Cell Culture and Methods Development
Cell culture technology transfer will begin with the expansion of a RCB to produce a WRCB and confirm media and procedures for viable cell density, doubling time and split ratios in small shaken cultures. During this activity, procedures for counting cells and expansion methods of the seed train for manufacturing will be established. The fed batch process will be confirmed in small shaken cultures to test the WRCB and fed batch culture process. Bench-top bioreactors will be used to confirm and finalize the fed batch process.
The culture method will be tested for scale up using our ATF Perfusion system. A Continuous Perfusion Bioreactor consisting of an ATF2 Perfusion system coupled to a 3L Applikon vessel is used to grow the seed culture up to 100 million cell density for seeding the 50L Continuous Perfusion Bioreactor (ATF6 Continuous Perfusion coupled to a 50L HyClone Single Use Bioreactor) controlled by GAMP5 Delta Software from Finesse. Material produced in this phase of the project will be used to purify the antibody protein using an AKTA Pilot under Continuous Purification mode (and for reference standard preparation and characterization.
Downstream process – Purification Methods
Harvest material produced above will be purified using an AKTA Pilot operating in Periodic Counter-current Chromatography (PCC) mode. Purified protein will be used to qualify existing lot release assays and to develop protein specific potency assays for including in the Phase I IND dossier. These small-scale purification runs will be done to confirm aspects of the continuous purification process such as minimum resin capacity, buffer volumes for wash/elution steps and product recovery.
The small scale purifications will provide background information for Straight-through Processing (STP) for polishing and viral inactivation step before proceeding to final UF/DF formulation and fill/finish. Key process intermediates will be frozen and tested for particles, SEC-HPLC and CIEF to confirm stability of frozen in-process intermediates intended for viral inactivation studies.