title: Efficiency improvement of an antibody production process by increasing the inoculum density.
abstract: Increasing economic pressure is the main driving force to enhance the efficiency of existing processes. We developed a perfusion strategy for a seed train reactor to generate a higher inoculum density for a subsequent fed batch production culture. A higher inoculum density can reduce culture duration without compromising product titers. Hence, a better capacity utilization can be achieved. The perfusion strategy was planned to be implemented in an existing large scale antibody production process. Therefore, facility and process constraints had to be considered. This article describes the initial development steps. Using a proprietary medium and a Chinese hamster ovary cell line expressing an IgG antibody, four different cell retention devices were compared in regard to retention efficiency and reliability. Two devices were selected for further process refinement, a centrifuge and an inclined gravitational settler. A concentrated feed medium was developed to meet facility constraints regarding maximum accumulated perfundate volume. A 2-day batch phase followed by 5 days of perfusion resulted in cell densities of 1.6 × 10(10) cells L(-1) , a 3.5 fold increase compared to batch cultivations. Two reactor volumes of concentrated feed medium were needed to achieve this goal. Eleven cultivations were carried out in bench and 50 L reactors showing acceptable reproducibility and ease of scale up. In addition, it was shown that at least three perfusion phases can be combined within a repeated perfusion strategy.
authors.affiliation: Pharmaceutical Biotechnology Div., Fraunhofer Inst. for Toxicology and Experimental Medicine (ITEM), Braunschweig, Germany.
authors.forename: Volker
authors.initials: V
authors.lastname: Hecht
authors.forename: Sevim
authors.initials: S
authors.lastname: Duvar
authors.forename: Holger
authors.initials: H
authors.lastname: Ziehr
authors.forename: Josef
authors.initials: J
authors.lastname: Burg
authors.forename: Alexander
authors.initials: A
authors.lastname: Jockwer
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D019149:Bioreactors; D016466:CHO Cells; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods; D003412:Cricetulus
chemical_list: D000911:Antibodies, Monoclonal
keywords: CHO; cell retention; inocululation enhancement; perfusion; seed train