Shear contributions to cell culture performance and product recovery in ATF and TFF perfusion systems.
Achievement of a robust and scalable cell retention device remains a challenge in perfusion systems. Of the two filtration systems commonly used, tangential flow filtration (TFF) systems often have an inferior product sieving profile compared to alternating tangential flow filtration (ATF) systems, which is typically attributed to the ATF's unique alternating flow. Here, we demonstrate that observed performance differences between the two systems are a function of cell lysis and not the alternating flow as previously thought. The peristaltic pump used in typical TFF perfusion systems is shown to be the single major contributor to shear stress and cell lysis. Replacing the peristaltic pump with a low shear centrifugal pump brought cell growth, cell lysis, particle concentration, and product sieving in a TFF perfusion system to levels comparable with that of an ATF. These results provide a correlation where poor product sieving can be partially explained by high shear in cell retention systems and demonstrate that low shear TFF systems are a feasible alternative to ATF systems.
Process Science, Boehringer Ingelheim, 4701 Kaiser Drive, Fremont, CA 94555, USA. Electronic address: samantha.wang@boehringer-ingelheim.com.
Samantha
S
Wang
Process Science, Boehringer Ingelheim, 4701 Kaiser Drive, Fremont, CA 94555, USA. Electronic address: scott.godfrey@boehringer-ingelheim.com.
Scott
S
Godfrey
Process Science, Boehringer Ingelheim, 4701 Kaiser Drive, Fremont, CA 94555, USA. Electronic address: janani.ravikrishnan.ext@boehringer-ingelheim.com.
Janani
J
Ravikrishnan
Process Science, Boehringer Ingelheim, 4701 Kaiser Drive, Fremont, CA 94555, USA. Electronic address: henry.lin@boehringer-ingelheim.com.
Henry
H
Lin
Process Science, Boehringer Ingelheim, 4701 Kaiser Drive, Fremont, CA 94555, USA. Electronic address: jens.vogel@boehringer-ingelheim.com.
Jens
J
Vogel
Process Science, Boehringer Ingelheim, 4701 Kaiser Drive, Fremont, CA 94555, USA. Electronic address: jon.coffman@boehringer-ingelheim.com.
Jon
J
Coffman
D061249:Batch Cell Culture Techniques / Q000379:methods*; D019149:Bioreactors; D002452:Cell Count; D049109:Cell Proliferation; D002470:Cell Survival; D005374:Filtration / Q000379:methods*
Alternating tangential flow; Cell lysis; Perfusion cell culture; Product sieving; Shear; Tangential flow filtration