Scaling up the downstream process for inactivated yellow fever vaccine: a pilot-scale proposal (Eukaryotes Pilot Laboratory/Bio-Manguinhos)

Introduction: Yellow Fever (YF) remains an endemic disease in tropical regions, with vaccination as the key element of prevention.To mitigate potential side effects associated with current live-attenuated YF 17DD vaccine, efforts are underway to develop an inactivated YF vaccine.This process, initially established at a bench scale, is now being scaled up to pilot production.Objectives: This study details the development and the proposal for implementation of a downstream purification process for 40L of viral suspension at pilot scale, enabling further process scaling and future clinical batch production. Methodology:The pilot-scale methodology was derived from experimental data obtained from benchscale processes, initially at 0.3L and subsequently at 1.5L, developed by the Laboratory of Physicochemical Analytical Technologies.The purification sequence included endonuclease digestion for host cell DNA degradation, followed by affinity and multimodal chromatography.Viral infectivity, residual DNA, and host cell protein (HCP) levels were rigorously monitored.Results: A GAP analysis, based on bench-scale data, identified critical factors for successful technical and operational adaptation to the 40L scale.Utilizing this analysis, a mass balance was developed to determine the inputs and outputs of each purification step, facilitating accurate volume estimations and alignment with production requirements.Simultaneously, supply chain planning ensured the availability of necessary materials and reagents.Standard operating procedures and production records (BPx) were established to guarantee traceability and consistency in the process.Water tests were conducted to validate the proposed process and optimize operational parameters for scale-up.A comprehensive process of control strategy is being implemented to standardize operations and ensure consistent product quality.The entire three-step purification process was meticulously mapped using a detailed flowchart created with the Bizagi tool. Conclusion:The results of YF virus downstream process at bench scale demonstrated the technical viability of producing IFA batches for an inactivated YF vaccine.This outcome, combined with scale-up studies, is crucial for project progression and future pilot-scale manufacturing.Leveraging Bio-Manguinhos' pilot plant infrastructure will facilitate project expansion and support upcoming clinical trials.