Premium
Efficiency improvement of an antibody production process by increasing the inoculum density
Author(s) -
Hecht Volker,
Duvar Sevim,
Ziehr Holger,
Burg Josef,
Jockwer Alexander
Publication year - 2014
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.1887
Subject(s) - perfusion , process engineering , scale up , bioreactor , pulp and paper industry , process (computing) , continuous production , biomedical engineering , chinese hamster ovary cell , centrifuge , chemistry , environmental science , computer science , chromatography , materials science , microbiology and biotechnology , cell culture , biology , medicine , physics , composite material , organic chemistry , classical mechanics , nuclear physics , engineering , cardiology , operating system , genetics
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. © 2014 American Institute of Chemical Engineers Biotechnol. Prog ., 30:607–615, 2014