title: Perfusion seed cultures improve biopharmaceutical fed-batch production capacity and product quality.
abstract: Volumetric productivity and product quality are two key performance indicators for any biopharmaceutical cell culture process. In this work, we showed proof-of-concept for improving both through the use of alternating tangential flow perfusion seed cultures coupled with high-seed fed-batch production cultures. First, we optimized the perfusion N-1 stage, the seed train bioreactor stage immediately prior to the production bioreactor stage, to minimize the consumption of perfusion media for one CHO cell line and then successfully applied the optimized perfusion process to a different CHO cell line. Exponential growth was observed throughout the N-1 duration, reaching >40 × 10(6) vc/mL at the end of the perfusion N-1 stage. The cultures were subsequently split into high-seed (10 × 10(6) vc/mL) fed-batch production cultures. This strategy significantly shortened the culture duration. The high-seed fed-batch production processes for cell lines A and B reached 5 g/L titer in 12 days, while their respective low-seed processes reached the same titer in 17 days. The shortened production culture duration potentially generates a 30% increase in manufacturing capacity while yielding comparable product quality. When perfusion N-1 and high-seed fed-batch production were applied to cell line C, higher levels of the active protein were obtained, compared to the low-seed process. This, combined with correspondingly lower levels of the inactive species, can enhance the overall process yield for the active species. Using three different CHO cell lines, we showed that perfusion seed cultures can optimize capacity utilization and improve process efficiency by increasing volumetric productivity while maintaining or improving product quality.
authors.affiliation: Cell Culture Development, Biogen Idec Inc., Research Triangle Park, NC, 27709.
authors.forename: William C
authors.initials: WC
authors.lastname: Yang
authors.forename: Jiuyi
authors.initials: J
authors.lastname: Lu
authors.forename: Chris
authors.initials: C
authors.lastname: Kwiatkowski
authors.forename: Hang
authors.initials: H
authors.lastname: Yuan
authors.forename: Rashmi
authors.initials: R
authors.lastname: Kshirsagar
authors.forename: Thomas
authors.initials: T
authors.lastname: Ryll
authors.forename: Yao-Ming
authors.initials: YM
authors.lastname: Huang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D061249:Batch Cell Culture Techniques; D019149:Bioreactors; D016466:CHO Cells; D003412:Cricetulus
chemical_list: D000911:Antibodies, Monoclonal
keywords: ATF; CHO cell culture; antibody; capacity; fed-batch; fusion protein; perfusion; product quality