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Aachen, Germany

Blesken C.,m2p-labs | Olfers T.,m2p-labs | Grimm A.,RWTH Aachen | Frische N.,m2p-labs
Engineering in Life Sciences | Year: 2016

The BioLector® Pro system from m2p-labs GmbH uses microtiter plates (MTPs) with an integrated microfluidic chip. By using microfluidic technology, the system can successfully carry out small-scale fed-batch cultivations. Working volumes of 0.8-1.5 mL are used to conduct cultivations as they were only as yet possible in lab fermenters. The measurements of biomass, fluorescence, pH and dissolved oxygen are performed by non-invasive optical methods. The control of pH and feeding rates are realized by micro-valves and micro-channels. For the first time, these unique microfluidic components achieve continuous feeding and pH control on an MTP format. Altogether, 32 bioreactor wells and 16 reservoir wells are placed on one plate. That means 32 fed-batch cultivations can be run in parallel, completely automated, with extensive data output. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim. Source

Huber R.,RWTH Aachen | Palmen T.G.,RWTH Aachen | Ryk N.,RWTH Aachen | Hillmer A.-K.,RWTH Aachen | And 3 more authors.
BMC Biotechnology | Year: 2010

Background: High-throughput cultivations in microtiter plates are the method of choice to express proteins from recombinant clone libraries. Such processes typically include several steps, whereby some of them are linked by replication steps: transformation, plating, colony picking, preculture, main culture and induction. In this study, the effects of conventional replication methods and replication tools (8-channel pipette, 96-pin replicators: steel replicator with fixed or spring-loaded pins, plastic replicator with fixed pins) on growth kinetics of Escherichia coli SCS1 pQE-30 pSE111 were observed. Growth was monitored with the BioLector, an on-line monitoring technique for microtiter plates. Furthermore, the influence of these effects on product formation of Escherichia coli pRhotHi-2-EcFbFP was investigated. Finally, a high-throughput cultivation process was simulated with Corynebacterium glutamicum pEKEx2-phoD-GFP, beginning at the colony picking step.Results: Applying different replication tools and methods for one single strain resulted in high time differences of growth of the slowest and fastest growing culture. The shortest time difference (0.3 h) was evaluated for the 96 cultures that were transferred with an 8-channel pipette from a thawed and mixed cryoculture and the longest time difference (6.9 h) for cultures that were transferred with a steel replicator with fixed pins from a frozen cryoculture. The on-line monitoring of a simulated high-throughput cultivation process revealed strong variances in growth kinetics and a twofold difference in product formation. Another experiment showed that varying growth kinetics, caused by varying initial biomass concentrations (OD600of 0.0125 to 0.2) led to strongly varying product formation upon induction at a defined point of time.Conclusions: To improve the reproducibility of high-throughput cultivation processes and the comparability between different applied cultures, it is strongly recommended to use automated or manual liquid handling stations or, alternatively, multi-channel pipettes. Because of their higher transfer volume and hence precision in comparison to pin replicators, they reduce the variance of initial biomass concentrations. With respect to the results obtained, other methods to increase the comparability between parallel cultivations by compensating differences in biomass concentrations are required, such as using autoinduction media, fed-batch operation of precultures or on-line monitoring in microtiter plates combined with automated liquid handling. © 2010 Huber et al; licensee BioMed Central Ltd. Source

Moussa M.,Minapharm Pharmaceuticals | Ibrahim M.,Minapharm Pharmaceuticals | El Ghazaly M.,Minapharm Pharmaceuticals | Rohde J.,Minapharm Pharmaceuticals | And 4 more authors.
BMC Biotechnology | Year: 2012

Background: Currently, the two most commonly used fibrinolytic agents in thrombolytic therapy are recombinant tissue plasminogen activator (rt-PA) and streptokinase (SK). Whereas SK has the advantage of substantially lower costs when compared to other agents, it is less effective than either rt-PA or related variants, has significant allergenic potential, lacks fibrin selectivity and causes transient hypotensive effects in high dosing schedules. Therefore, development of an alternative fibrinolytic agent having superior efficacy to SK, approaching that of rt-PA, together with a similar or enhanced safety profile and advantageous cost-benefit ratio, would be of substantial importance. Pre-clinical data suggest that the novel fibrinolytic recombinant staphylokinase (rSAK), or related rSAK variants, could be candidates for such development. However, since an efficient expression system for rSAK is still lacking, it has not yet been fully developed or evaluated for clinical purposes. This study's goal was development of an efficient fermentation process for the production of a modified, non-glycosylated, biologically active rSAK, namely rSAK-2, using the well-established single cell yeast Hansenula polymorpha expression system.Results: The development of an efficient large scale (80 L) Hansenula polymorpha fermentation process of short duration for rSAK-2 production is described. It evolved from an initial 1mL HTP methodology by successive scale-up over almost 5 orders of magnitude and improvement steps, including the optimization of critical process parameters (e.g. temperature, pH, feeding strategy, medium composition, etc.). Potential glycosylation of rSAK-2 was successfully suppressed through amino acid substitution within its only N-acetyl glycosylation motif. Expression at high yields (≥ 1g rSAK-2/L cell culture broth) of biologically active rSAK-2 of expected molecular weight was achieved.Conclusion: The optimized production process described for rSAK-2 in Hansenula polymorpha provides an excellent, economically superior, manufacturing platform for a promising therapeutic fibrinolytic agent. © 2012 Moussa et al.; licensee BioMed Central Ltd. Source

Kottmeier K.,RWTH Aachen | Kottmeier K.,TU Dresden | Muller C.,RWTH Aachen | Muller C.,m2p-labs | And 2 more authors.
Applied Microbiology and Biotechnology | Year: 2010

By the use of directed limitations of secondary substrates, the metabolic flux should be deflected from biomass production to product formation. In order to study the impact of directed limitations caused by various secondary substrates on the growth and product formation of the methylotrophic yeast Hansenula polymorpha, the cultivation systems respiration activity monitoring system (RAMOS) and BioLector were used in parallel. While the RAMOS device allows the online monitoring of the oxygen transfer rate in shake flasks, the BioLector enables in microtiter plates the monitoring of scattered light and the fluorescence intensity of the green fluorescent protein (GFP). Secondary substrate limitations of phosphate, potassium, and magnesium were analyzed in batch fermentations. The sole carbon source was either 10 g/L glucose or 10 g/L glycerol. The expression of the GFP gene is controlled by the FMD promoter (formate dehydrogenase). In batch cultures with glucose as carbon source, a directed limitation of phosphate increased the GFP production 1.87-fold, compared to phosphate unlimited conditions. Under potassium-limited conditions with glycerol as sole carbon source, the GFP production was 1.41-fold higher compared to unlimited conditions. A limitation of the substrate magnesium resulted in a 1.22-fold increase GFP formation in the case of glycerol as carbon source. © 2009 Springer-Verlag. Source

News Article | November 7, 2013
Site: www.finsmes.com

The round was led by new investor Fidura Private Equity Fonds (which acquired a total stake of 27.61% in the company), with participation from High-Tech-Gründerfonds. The company intends to use the funds to further expand its technology with new developments and product improvements, particularly in the field of microscale process control (fed-batch and pH control) and automation of its BioLector®, and to increase its global sales network, especially in the USA and Asia. Co-founded in 2005 by and Frank Kensy and Carsten Müller, m2p-labs (“m2p” stands for “from microreactor to process”) is engaged in the development and sales of lab analysis systems (microbioreactors) to carry out high throughput test series in the field of cellular screening and bioprocess development. The company focuses on microreaction and automated solutions for screening and bioprocess development. Commercialized products include the BioLector® and the FlowerPlate®, which provide an intelligent micro fermentation platform to enable the biotechnology, chemical and pharmaceutical industry to increase their number and information of microbial and cell culture experiments. Customers include pharmaceutical and chemical companies, universities (ETH Zürich, RWTH Aachen, University College London and Imperial College London) and research institutions such as the Jülich Research Centre.

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