Improving the batch-to-batch reproducibility in microbial cultures during recombinant protein production by guiding the process along a predefined total biomass profile (original) (raw)
Related papers
Journal of Biotechnology, 2007
Batch-to-batch reproducibility of fermentation processes performed during the manufacturing processes of biologics can be increased by operating the cultures at feed rate profiles that are robust against typically arising disturbances. Remaining randomly appearing deviations from the desired path should be suppressed automatically by manipulating the feed rate. With respect to the cells' physiology it is best guiding the cultivations along an optimal profile of the specific biomass growth rate μ(t). However, there are two problems that speak for further investigations: Upon severe disturbances that may happen during the fermentation, the biomass concentration X may significantly deviate from its desired value, then a fixed μ-profile leads to a diminished batch-tobatch reproducibility. Second, the specific growth rate cannot easily be estimated online to a favourably high accuracy, hence it is difficult to determine the deviations in μ from the desired profile. The alternative discussed here solves both problems by keeping the process at the corresponding total cumulative carbon dioxide production-profile: it is robust against distortions in X and the controlled variable can accurately be measured online during cultivations of all relevant sizes. As compared to the fermentation practice currently used in industry, the experimental results, presented at the example of a recombinant protein production with Escherichia coli cells, show that CPR-based corrections lead to a considerably improved batch-to-batch reproducibility.
Advanced monitoring & control in microbial cultivation processes for recombinant protein production
2018
Verglichen mit dem rasanten Fortschreiten in den molekularbiologischen Wissenschaften während der letzten Jahrzehnte, konnten die Entwicklungen, der in der industriellen Praxis verwendeten Fermentations- und Downstream-Techniken, bei weitem nicht mithalten. Daraus ergibt sich ein Ungleichgewicht zwischen neuen biologischen Systemen und den industriellen Produktionstechnologien, was zu einer fortwährenden Abnahme der jährlichen Neuanmeldungen von Produktionsprozessen für Biologics führt. In ihrer PAT-Initiative (PAT = Process Analytical Technology) benennt die für die Zulassung von Pharmaprodukten zuständige US-amerikanische Aufsichtsbehörde FDA die Problempunkte, welche verbessert werden müssen, und fordert die Hersteller gleichzeitig auf in wissenschaftlich basierte Ansätze zur Problemlösung zu investieren. Diese Doktorarbeit zeigt am Beispiel der rekombinanten Proteinproduktion mit Escherichia coli - Bakterien verschiedene bioverfahrenstechnische Ansätze, wie diesen Forderungen na...
Microbial Cell Factories, 2009
The optimisation and scale-up of process conditions leading to high yields of recombinant proteins is an enduring bottleneck in the post-genomic sciences. Typical experiments rely on varying selected parameters through repeated rounds of trial-and-error optimisation. To rationalise this, several groups have recently adopted the 'design of experiments' (DoE) approach frequently used in industry. Studies have focused on parameters such as medium composition, nutrient feed rates and induction of expression in shake flasks or bioreactors, as well as oxygen transfer rates in micro-well plates. In this study we wanted to generate a predictive model that described small-scale screens and to test its scalability to bioreactors.
Journal of Visualized Experiments
A core business in industrial biotechnology using microbial production cell factories is the iterative process of strain engineering and optimization of bioprocess conditions. One important aspect is the improvement of cultivation medium to provide an optimal environment for microbial formation of the product of interest. It is well accepted that the media composition can dramatically influence overall bioprocess performance. Nutrition medium optimization is known to improve recombinant protein production with microbial systems and thus, this is a rewarding step in bioprocess development. However, very often standard media recipes are taken from literature, since tailor-made design of the cultivation medium is a tedious task that demands microbioreactor technology for sufficient cultivation throughput, fast product analytics, as well as support by lab robotics to enable reliability in liquid handling steps. Furthermore, advanced mathematical methods are required for rationally analyzing measurement data and efficiently designing parallel experiments such as to achieve optimal information content. The generic nature of the presented protocol allows for easy adaption to different lab equipment, other expression hosts, and target proteins of interest, as well as further bioprocess parameters. Moreover, other optimization objectives like protein production rate, specific yield, or product quality can be chosen to fit the scope of other optimization studies. The applied Kriging Toolbox (KriKit) is a general tool for Design of Experiments (DOE) that contributes to improved holistic bioprocess optimization. It also supports multi-objective optimization which can be important in optimizing both upstream and downstream processes. Video Link The video component of this article can be found at https://www.jove.com/video/56234/ Nutrition medium optimization is known to improve recombinant protein production with microbial systems 14,15,16,17 and consequently, the adjustment of medium composition is a rewarding step in bioprocess development with respect to optimal productivity 18,19,20,21. Intense research on the application of microtiter plates (MTPs) for microbial cultivation 22,23,24 paved the way for the development and design of MTPs for microbial cultivation 25,26 and the development of MTP-based microbioreactor (MBR) systems with online monitoring and environmental control 27,28
Impact of high throughput technology on recombinant protein production
2006
... Bullock Jonathan ; Hudson Ian ; Hall Richard ; Jenkins Owen ; Cole Jeffrey. Abstract: Journal: Microbial Cell Factories. Issn: 14752859. EIssn: Year: 2006. Volume: 5. Issue: Suppl+1. pages/rec.No: S30. Keywords, ... DOAJ - Directory of Open Access Journals | 2011 | Lund University ...