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.