Wide‐surface pore microfiltration membrane drastically improves sieving decay in TFF‐based perfusion cell culture and streamline chromatography integration for continuous bioprocessing

Abstract

Although several compelling benefits for bioprocess intensification have been reported, the need for a streamlined integration of perfusion cultures with capture chromatography still remains unmet. Here, a robust solution is established by conducting tangential flow filtration-based perfusion with a wide-surface pore microfiltration membrane. The resulting integrated continuous bioprocess demonstrated negligible retention of antibody, DNA, and host cell proteins in the bioreactor with average sieving coefficients of 98 ± 1%, 124 ± 28%, and 109 ± 27%, respectively. Further discussion regarding the potential membrane fouling mechanisms is also provided by comparing two membranes with different surface pore structures and the same hollow fiber length, total membrane area, and chemistry. A cake-growth profile is reported for the narrower surface pore, 0.65-µm nominal retention perfusion membrane with final antibody sieving coefficients ≤70%. Whereas the sieving coefficient remained ≥85% during 40 culture days for the wide-surface pore, 0.2-µm nominal retention rating membrane. The wide-surface pore structure, confirmed by scanning electron microscopy imaging, minimizes the formation of biomass deposits on the membrane surface and drastically improves product sieving. This study not only offers a robust alternative for integrated continuous bioprocess by eliminating additional filtration steps while overcoming sieving decay, but also provides insight into membranes' fouling mechanism.

Modeling flux in tangential flow filtration using a reverse asymmetric membrane for Chinese hamster ovary cell clarification

“…In a recent study, Pinto and Brower 25 investigated the performance of the UJP and UMP microfilters for clarification during perfusion operations. They observed that the sieving coefficient for the smaller nominal pore size UMP microfilter was superior to the larger nominal pore size UJP microfilter.…”

An innovative strategy to recycle permeate in biologics continuous manufacturing process to improve material efficiency and sustainability

“…Fully closed processing relied on using only peristaltic pumps and having the entire flow path being either gamma irradiated or autoclaved, including tubing, multi‐column BioSMB (Sartorius AG, Goettingen, Germany) manifold and Protein A columns. Aseptic techniques were employed for connections through tube welding with single use thermoplastic tubing (Pharmed L/S 16, Masterflex, PA) 18–20 . ProA FT was further processed via two different methods.…”

A common framework for integrated and continuous biomanufacturing

“…Performance comparison of ATF to TFF show a lower sieving decay for ATF, potentially due to the bidirectional flow, minimizing the biomaterial deposit on the hollow fiber membrane (Radoniqi et al, 2018; Wang et al, 2017). However the ATF has been associated with operational instability at higher cell densities (Pinto & Brower, 2020; Pinto et al 2020). Backflushing of the ATF and TFF filters with perfusate or fresh media has shown potential to extend filtration life during a manufacturing campaign (Wang et al, 2017; Wang et al, 2019).…”