Biomedical Engineering and Bioengineering Commons^™

Integrated Continuous Bioprocessing – Opportunities And Challenges, Mats Akesson, Martin Heitmann, Ditte Skipstrup, Peter Tiainen

Integrated Continuous Biomanufacturing II

We have developed an integrated continuous processing framework for end-to-end production of complex fragile proteins based on perfusion cultivation and automated multi-step purification [1,2,3]. Upstream, the integrated system consists of a stirred tank bioreactor with an ATF cell retention system. The clarified harvest directly enters an off-the-shelf ÄKTA chromatography system converted into a continuous purification unit. Two alternating capture columns precede an automated multi-step purification train with full flexibility and control of individual columns.

The integrated set-up enables compact automated bench-top factories converting cell culture media to purified protein in an efficient manner without intermediate storage. It provides monitoring of …

Bhk Cells Physiological Response To Spin-Filter Stress Condition, Bruno Labate, Aldo Tonso

Integrated Continuous Biomanufacturing II

Mammalian cells are the preferable platform for secreted pharmaceutical proteins due to the compatibility of posttranslational modifications performed by these cells with therapeutic applications. However, to obtain a desirable quantity of these proteins, high cell concentrations are also needed. Baby hamster kidney BHK-21/C13 cells, adapted to single-cell suspension culture growth were cultivated in batch mode in a Biostat B (Sartorius AG, Germany) bioreactor with a 1,5 L working volume vessel. The temperature was controlled at 37 ºC, pH at 7.2 with CO­₂, agitation at 80 rpm and dissolved oxygen at 50% of air saturation. The culture medium used …

Monitoring Intracellular Component Pools To Identify Steady State In Mammalian Cell Perfusion Culture, Daniel Karst, Robert Steinhoff, Marie Kopp, Renato Zenobi, Massimo Morbidelli, Miroslav Soos

Integrated Continuous Biomanufacturing II

Perfusion cultures of mammalian cells are a well-considered alternative in the process development for new biologic products. Stable operation of the culture should eventually lead to a steady state, positively effecting product quality. Although macroscopic variables such as viable cell density, main metabolite or product concentrations are observed constant, little is known about the extent of steady state on intracellular level.

In this study intracellular component pools of CHO cells were monitored at different steady states in perfusion cultures using matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). A novel single extraction was used to separate metabolite, …