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.

No abstract

There are few reports of the adoption of continuous processes in bioproduction, particularly the implementation of end‐to‐end continuous or integrated processes, due to difficulties such as feed adjustment and incorporating virus filtration. Here, we propose an end‐to‐end integrated 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 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 defines 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 (liters per effective surface area of filter in square meters per hour) confirmed robust virus reduction over these ranges. Complete clearance with virus logarithmic reduction value ≥4 was achieved even with a process pause at the lowest flux. The end‐to‐end integrated continuous process proposed in this study is amenable to production processes, and the investigated virus filters have excellent applicability to continuous processes conducted at constant flux.

Multi-column chromatography (periodic counter-current chromatography, PCCC) with protein A affinity resins is used as an efficient monoclonal antibody (mAb) continuous capture process. However, the evaluation of the efficiency of PCCC process compared with repeated batch (or cyclic) operation (RBO) is still not standardized. In this study, we proposed two criteria for evaluating the capture process efficiency, productivity P and buffer consumption per load amount, BC. These criteria were calculated for 2 column, 3 column and 4 column-PCCC and for RBO. For PCCC, P was proportional to mAb concentration C0 and inversely proportional to the number of columns, nC since the loaded amount of mAb ML was fixed for the calculation. Bc did not depend on C0 or nC as ML was the same. For RBO, BC was larger by 20% at residence time (RT) = 4 minutes, and by 50% at RT = 2 minutes whereas P was similar to or slightly lower than that for 2C-PCCC at the same RT (4 minutes).