Viral clearance in end-to-end continuous process for mAb purification: Total flow-through integrated polishing on two columns connected to virus filtration

Abstract

There are few reports of the adoption of continuous processes in bioproduction, particularly the implementation of end-to-end continuous processes, due to difficulties such as feed adjustment, production batch demarcation, and incorporating virus filtration. Here, we propose an end-to-end continuous process for a monoclonal antibody (mAb) with three integrated process segments: upstream production processes with pool-less direct connection, pooled low pH virus inactivation with automated pH control and a total flow-through integrated polishing process in which two columns were directly connected with a virus filter. The pooled virus inactivation step demarcates the batch, and high impurities reduction and mAb recovery were achieved for batches conducted in succession. Viral clearance tests also confirmed robust virus reduction for the flow-through two column chromatography and the virus filtration steps. Additionally, viral clearance tests with two different hollow fiber virus filters operated at flux ranging from 1.5 to 40 LMH confirmed robust virus reduction over these ranges. Complete clearance with LRV ≥ 4 was achieved even with a process pause at the lowest flux. The end-to-end continuous process proposed in this study is highly applicable to production processes, and the investigated virus filters have excellent applicability to continuous processes conducted at constant flux.

Integrated continuous downstream process of monoclonal antibody developed by converting the batch platform process based on the process characterization

“…The detailed analysis of the two‐connected FTC column operation with VF attached to the 2nd column including viral clearance studies has been reported in Shirataki et al (2023).…”

“…The recovery of Run 7 was lower due to the misoperations (some of the pool was lost). The detailed analysis on Run 4 was published in Shirataki et al (2023).…”

Residence time distribution in continuous virus filtration

“…A fully end‐to‐end continuous biomanufacturing process must include continuous virus filtration (David et al, 2019). Research on continuous virus filtration processes is still in early stages (Bohonak et al, 2021; Fan et al, 2021; Lute et al, 2020; Shirataki et al, 2023). It is crucial to elucidate the differences between batch and continuous operations, and to master the methods for designing operation parameters of continuous processes based on batch operations.…”

“…Establishing robust and efficient models require proper model calibration and suitable calibration experiments (Chen et al, 2022; Chen, Chen, et al, 2023; Chen, Yao, et al, 2023; Yang et al, 2024). In virus filtration, pressure operations have been primarily applied and modeled, rather than considering constant flux processes (Shirataki et al, 2023, 2021). Recently, Malakian et al (2022) developed a PF‐CST‐CST model to describe the RTD of virus filtration in constant flux processes.…”