Microbioreactor System Research Articles

Parallel Multifactorial Process Optimization and Intensification for High-Yield Production of Live YF17D-Vectored Zika Vaccine.

The live-attenuated yellow fever 17D strain is a potent vaccine and viral vector. Its manufacture is based on embryonated chicken eggs or adherent Vero cells. Both processes are unsuitable for rapid and scalable supply. Here, we introduce a high-throughput workflow to identify suspension cells that are fit for the high-yield production of live YF17D-based vaccines in an intensified upstream process. The use of an automated parallel ambr15 microbioreactor system for screening and process optimization has led to the identification of two promising cell lines (AGE1.CR.pIX and HEKDyn) and the establishment of optimized production conditions, which have resulted in a >100-fold increase in virus titers compared to the current state of the art using adherent Vero cells. The process can readily be scaled up from the microbioreactor scale (15 mL) to 1 L stirred tank bioreactors. The viruses produced are genetically stable and maintain their favorable safety and immunogenicity profile, as demonstrated by the absence of neurovirulence in suckling BALB/c mice and consistent seroprotection in AG129 mice. In conclusion, the presented workflow allows for the rapid establishment of a robust, scalable, and high-yield process for the production of live-attenuated orthoflavivirus vaccines, which outperforms current standards. The approach described here can serve as a model for the development of scalable processes and the optimization of yields for other virus-based vaccines that face challenges in meeting growing demands.

Revisiting the urease production of MICP-relevant bacterium Sporosarcina pasteurii during cultivation

The ureolytic bacterium Sporosarcina pasteurii is commonly used for Microbial Induced Calcium Carbonate Precipitation (MICP). This process has a variety of applications in the fields of construction, geotechnical engineering, and environmental remediation. However, the factors influencing urease activity of S. pasteurii during cultivation are not yet fully understood. Even though there is debate over whether higher urease activity is in fact beneficial for MICP applications, knowledge about urease production can help provide ureolytic biomass more cost-effectively, reducing overall production costs. This study revisits the effect of various cultivation conditions on growth and urease activity by using an automated high-throughput microbioreactor system combined with a novel system for high-throughput urease activity quantification. This experimental set-up allows for unprecedented data density and enables new insights into urease production of S. pasteurii.Several factors were tested, including oxygen limitation, the feeding of urea, choice of ammonium salt, the proportion of ammonium salt in the medium, and lowering the pH during cultivation; none of these had a notable impact on urease production. Only the addition of a broad spectrum of nutrients through regular feeding with highly concentrated yeast extract and glucose led to a 70 % increased specific urease activity compared to a batch culture.These results can improve the understanding of the regulatory mechanisms governing urease expression in S. pasteurii during cultivation, and allow to adapt the cultivation process accordingly. Additionally, the new system for automated high-throughput enzyme activity determination presented in this study could be applied to optimize bioprocesses involving other ion producing enzymes.

Optimization of a Bacterial Cultivation Medium via a Design-of-Experiment Approach in a Sartorius Ambr® 15 Fermentation Microbioreactor System

In the evolving landscape of sustainable biopharmaceutical process development, the utilization of bacteria in the production of various compounds via fermentation has attracted extensive attention from scientists. A successful fermentation process and the release of its associated products hinge on the synergy between an efficient bacterial strain and the formulation of a suitable growth medium. Balancing all nutrient levels of a growth medium to maximize microbial growth and the product quality is quite an intricate task. In this context, significant advancements have been achieved via the strategic implementation of design-of-experiment (DOE) methodologies and the utilization of parallel microbioreactor systems. This work presents a case study of the fermentation growth medium optimization of a Gram-negative bacterium of the Neisseriaceae family that releases outer membrane vesicles (OMVs), which represent a potential vaccine platform. To achieve this, the ability of Sartorius MODDE®13 DOE software to explore multiple variables and their interactions was combined with the functionality of a Sartorius Ambr® 15F parallel microbioreactor system. The findings reported in this study have led to the design of a well-suited fermentation medium for a Gram-negative bacterium and an improvement in the quality of the OMVs produced from it.

Bioprospecting of Actinobacterial Diversity and Antibacterial Secondary Metabolites from the Sediments of Four Saline Lakes on the Northern Tibetan Plateau.

The Tibetan Plateau, known as the "Roof of the World" and "The Third Pole", harbors numerous saline lakes primarily distributed in the Northern Tibetan Plateau. However, the challenging conditions of high altitude, low oxygen level, and harsh climate have limited investigations into the actinobacteria from these saline lakes. This study focuses on investigating the biodiversity and bioactive secondary metabolites of cultivable actinobacteria isolated from the sediments of four saline lakes on the Northern Tibetan Plateau. A total of 255 actinobacterial strains affiliated with 21 genera in 12 families of 7 orders were recovered by using the pure culture technique and 16S rRNA gene phylogenetic analysis. To facilitate a high-throughput bioactivity evaluation, 192 isolates underwent OSMAC cultivation in a miniaturized 24-well microbioreactor system (MATRIX cultivation). The antibacterial activity of crude extracts was then evaluated in a 96-well plate antibacterial assay. Forty-six strains demonstrated antagonistic effects against at least one tested pathogen, and their underlying antibacterial mechanisms were further investigated through a dual-fluorescent reporter assay (pDualrep2). Two Streptomyces strains (378 and 549) that produce compounds triggering DNA damage were prioritized for subsequent chemical investigations. Metabolomics profiling involving HPLC-UV/vis, UPLC-QTOF-MS/MS, and molecular networking identified three types of bioactive metabolites belonging to the aromatic polyketide family, i.e., cosmomycin, kidamycin, and hedamycin. In-depth analysis of the metabolomic data unveiled some potentially novel anthracycline compounds. A genome mining study based on the whole-genome sequences of strains 378 and 549 identified gene clusters potentially responsible for cosmomycin and kidamycin biosynthesis. This work highlights the effectiveness of combining metabolomic and genomic approaches to rapidly identify bioactive chemicals within microbial extracts. The saline lakes on the Northern Tibetan Plateau present prospective sources for discovering novel actinobacteria and biologically active compounds.

LED Illumination Modules Enable Automated Photoautotrophic Cultivation of Microalgae in Parallel Milliliter-Scale Stirred-Tank Bioreactors

Scalable lab-scale photobioreactors are needed for the exploration of new and improved photoautotrophic bioprocesses. Microbioreactor systems in which parallel bioreactors operate automatically are frequently employed to increase the speed of strain selection as well as the bioprocess-based exploration of heterotrophic fermentation processes. To enable the photoautotrophic operation of a commercially available parallel microbioreactor system with 48 stirred-tank bioreactors, LED illumination modules were designed to allow for individual light supply (400–700 nm) for each of the parallel bioreactors automated by a liquid handling station that performs both individual pH control and OD750 detection. The illumination modules enable dynamic variation of the incident light intensities of up to 1800 µmol m−2 s−1. Automated liquid level detection and volume control of each individual mL-scale gassed photobioreactor has to be established to compensate for evaporation because of the long process times of several days up to weeks. Photoautotrophic batch processes with Microchloropsis salina that employ either varying constant incident light intensities or day and night dynamics resulted in a standard deviation of OD750 of up to a maximum of 10%, with the exception of high-photoinhibiting incident light intensities. The established photoautotrophic microbioreactor system enables the automated investigation of microalgae processes in up to 48 parallel stirred photobioreactors and is thus a new tool that enables efficient characterization and development of photoautotrophic processes with microalgae.

Modeling of a capillary microbioreactor for VOC removal

Capillary microreactors have shown superior performance as a result of the precise control of reaction parameters, contact time, shape and size of the interface between gas and liquid and enhanced mass transfer due to high surface to volume ratio. In this work, a mathematical model of a capillary microbioreactor treating VOC vapors was developed and validated experimentally. The main goal for this conceptual model was to adequately describe or predict VOC vapor removal in the capillary microbioreactor under different operating conditions and its mineralization to CO2. The model considered toluene as the model VOC and included oxygen consumption and carbon dioxide production. Experimental data used for model calibration and validation were obtained from experiments conducted at various toluene and volumetric loading rates. A parametric sensitivity to the model was performed to identify the parameters that most affected the performance of the microbioreactor system. The most sensitive parameters were biokinetic parameters followed by system specific parameters, indicating that the system performance was limited by kinetics rather than by mass transfer. Overall, the model and the approach described herein help us better understand the performance and limits of these microbioreactors and they serve as important tools for process optimization.

Cultivation of Mycolicibacterium spp. Mutants in Miniaturized and High-Throughput Format to Characterize Their Growth, Phytosterol Conversion Ability, and Resistance to the Steroid Products.

This chapter describes methods for cultivation and characterization of the growth of Mycolicibacterium spp. mutants in a microbioreactor system in the presence of steroids and/or phytosterols followed by high-throughput mass spectrometry analysis to describe their ability to convert phytosterols into the target steroid androstenedione (AD). We focus on Mycolicibacterium neoaurum NRRL B-3805 ΔkstD which can convert phytosterol into androstenedione (AD) as one of its major steroid products, and mutants thereof with increased tolerance towards this end-product. By using BioLector 48-well plates with optodes at the bottom of each well, bacterial growth can be monitored online despite the turbidity of the growth medium resulting from non-dissolved phytosterol and steroid particles. To cope with the large number of samples that accumulate during growth experiments in microbioreactors and similar formats (e.g., microtiter plates), protocols for extraction and subsequent RapidFire-MS analysis are presented. This reduces the analysis time per sample to 10 sfrom 10min required forregular LC-MS analysis.

Stirred culture of cartilaginous microtissues promotes chondrogenic hypertrophy through exposure to intermittent shear stress.

Cartilage microtissues are promising tissue modules for bottom up biofabrication of implants leading to bone defect regeneration. Hitherto, most of the protocols for the development of these cartilaginous microtissues have been carried out in static setups, however, for achieving higher scales, dynamic process needs to be investigated. In the present study, we explored the impact of suspension culture on the cartilage microtissues in a novel stirred microbioreactor system. To study the effect of the process shear stress, experiments with three different impeller velocities were carried out. Moreover, we used mathematical modeling to estimate the magnitude of shear stress on the individual microtissues during dynamic culture. Identification of appropriate mixing intensity allowed dynamic bioreactor culture of the microtissues for up to 14 days maintaining microtissue suspension. Dynamic culture did not affect microtissue viability, although lower proliferation was observed as opposed to the statically cultured ones. However, when assessing cell differentiation, gene expression values showed significant upregulation of both Indian Hedgehog (IHH) and collagen type X (COLX), well known markers of chondrogenic hypertrophy, for the dynamically cultured microtissues. Exometabolomics analysis revealed similarly distinct metabolic profiles between static and dynamic conditions. Dynamic cultured microtissues showed a higher glycolytic profile compared with the statically cultured ones while several amino acids such as proline and aspartate exhibited significant differences. Furthermore, in vivo implantations proved that microtissues cultured in dynamic conditions are functional and able to undergo endochondral ossification. Our work demonstrated a suspension differentiation process for the production of cartilaginous microtissues, revealing that shear stress resulted to an acceleration of differentiation towards hypertrophic cartilage.

Developing a fluorometric urease activity microplate assay suitable for automated microbioreactor experiments.

Quantifying urease activity is an important task for Microbial Induced Calcite Precipitation research. A new urease activity microplate assay using a fluorescent pH indicator is presented. The method is also suitable for automated measurements during microbioreactor experiments. The assay reagent consists of the green fluorescent pH-indicator fluorescein, urea and a phosphate buffer. After sample addition, the microbial urease hydrolyses urea, which results in a pH and hence fluorescence increase. The fluorescence signal can be measured with a microplate reader or with the microbioreactor system BioLector, allowing for automated urease activity measurements during cultivation experiments. In both measurement systems, the fluorescence signal slope highly correlates with the urease activity measured offline with standard methods. Automated measurement is possible, as no sample preparation such as centrifugation or adjusting of the optical density is required. The assay was developed so that the culture samples turbidity, salinity or buffer concentration does not have a negative impact on the fluorescence signal. The assay allows for straightforward, non-hazardous, parallelized, cheap and reliable measurements, making research on ureolytic bacteria for Microbial Induced Calcite Precipitation more efficient. The assay could be adapted to other enzymes, which have a strong impact on the pH value.

Definitive screening accelerates Taxol biosynthetic pathway optimization and scale up in Saccharomyces cerevisiae cell factories.

Recent technological advancements in synthetic and systems biology have enabled the construction of microbial cell factories expressing diverse heterologous pathways in unprecedentedly short time scales. However, the translation of such laboratory scale breakthroughs to industrial bioprocesses remains a major bottleneck. In this study, an accelerated bioprocess development approach was employed to optimize the biosynthetic pathway of the blockbuster chemotherapy drug, Taxol. Statistical design of experiments approaches were coupled with an industrially relevant high-throughput microbioreactor system to optimize production of key Taxol intermediates, Taxadien-5α-ol and Taxadien-5α-yl-acetate, in engineered yeast cell factories. The optimal factor combination was determined via data driven statistical modelling and validated in 1 L bioreactors leading to a 2.1-fold improvement in taxane production compared to a typical defined media. Elucidation and mitigation of nutrient limitation enhanced product titers a further two-fold and titers of the critical Taxol precursors, Taxadien-5α-ol and Taxadien-5α-yl-acetate were improved to 34 and 11mg L-1 , representing a three-fold improvement compared to the highest literature titers in S. cerevisiae. Comparable titers were obtained when the process was scaled up a further five-fold using 5 L bioreactors. The results of this study highlight the benefits of a holistic design of experiments guided approach to expedite early stage bioprocess development.

Evaluation of a microbioreactor system as a screening tool for optimizing lentiviral vector process development in suspension culture

With an increasing number of lentiviral vector (LVV)-based cell and gene therapy candidates reaching clinical trials, scalable suspension cell culture processes using stirred tank reactors (STRs) are needed to meet future demands. However, to cost-effectively scale LVV production in STRs requires process development which can be expensive and time consuming to perform in bench-top bioreactors. To address these issues, a multi-parametric approach for process development using a micro scale bioreactor system (Ambr® 15 cell culture system, Sartorius) was assessed. Since the medium exchange process step cannot be linearly or methodically scaled-down from a bench-scale STR to a microbioreactor due to system differences, this study focused on adjusting to those differences by developing and testing three different medium exchange protocols. The implementation of one approach (Process 2.0) using an automated cell settling medium exchange protocol produced results which closely aligned with an established LVV bench-scale process in transfection efficiency and productivity, as well as lowered variability between vessels in the cell culture workstation. In summary, this study demonstrates the suitability of the Ambr 15® system as a process screening tool which has the potential to reduce costs and timelines of the development of scalable LVV production systems in suspension culture.

Equal mixing time enables scale‐down and optimization of a CHO cell culture process using a shaken microbioreactor system

The advancement of microbioreactor technology in recent years has transformed early- and mid-stage process development. The monitoring and control capabilities of microbioreactors not only promote the quick accumulation of process knowledge but has also led to an increased scalability when compared to traditionally used systems such as shake flasks and microtitre plates. This study seeks to establish a framework for the micro-Matrix microbioreactor (Applikon-Biotechnology BV) as process development tool. Using the Dual Indicator System for Mixing Time, the system was initially characterized for mixing properties at varying operating conditions, which was found to yield mixing times between 0.9 and 41.8 s. A matched mixing time was proposed as scale-down criterion for an IgG4 producing GS-CHO fed-batch process between a 5 L stirred tank reactor (STR) and the micro-Matrix microbioreactor. Growth trends, maximum viable cell concentrations, final titre, and glycoprofiles were nearly identical at both scales. The scale-down model was then employed to optimize a bolus feeding regime using response surface methodology, which led to a 25.4% increase of the space-time yield and a 25% increase of the final titre. The optimized feeding strategy was validated at the small-scale and successfully scaled up to the 5 L STR. This work for the first time provides a framework of how the micro-Matrix microbioreactor can be implemented in a bioprocess development workflow and demonstrates scalability of growth and production kinetics as well as IgG4 glycosylation between the micro-Matrix and a benchtop-scale STR system.

Https://www.insights.bio/cell-and-gene-therapy-insights/journal/article/2128/Scalable-upstream-process-development-for-the-suspension-based-production-of-lentiviral-vectors-for-CAR-T-cell-therapies-with-multiparallel-benchtop

Cell and gene-based therapies present a new treatment paradigm that have the potential to address unmet clinical needs. Viral vectors such as adenoviruses, adeno-associated viruses, retroviruses and lentiviruses are effective delivery systems for genetic material used in cell and gene therapies and vaccines. HEK-293 cells and derivatives are commonly used as a workhorse cell line for lentiviral vector (LVV) production for cell and gene therapy applications. Adherent production processes with these cells utilize static flask cultures, and this adherent method is quite easy to develop and perform. However, it also significantly lacks the ability for automation and scalability. Typical bioreactors based either on a rocking motion or stirred tank agitation can provide these features. Therefore, to scale up viral vector production for commercialization, adherent processes should be shifted to a suspension-based process, a significant challenge for the regenerative medicine industry. The use of a suspension adapted HEK-293 cell line and the Ambr^{^® }15 microbioreactor system can facilitate transition from adherent cultures to suspension cultures by enabling fast process optimization with the ability to screen in parallel many parameters in small volumes. As a proof-of-concept study, we established here such a transitional protocol for the cultivation of suspension adapted HEK-293T cells and the production of CD19-CAR lentivirus in small and benchtop scale stirred bioreactors.

High-throughput microbioreactor provides a capable tool for early stage bioprocess development

Tremendous advancements in cell and protein engineering methodologies and bioinformatics have led to a vast increase in bacterial production clones and recombinant protein variants to be screened and evaluated. Consequently, an urgent need exists for efficient high-throughput (HTP) screening approaches to improve the efficiency in early process development as a basis to speed-up all subsequent steps in the course of process design and engineering. In this study, we selected the BioLector micro-bioreactor (µ-bioreactor) system as an HTP cultivation platform to screen E. coli expression clones producing representative protein candidates for biopharmaceutical applications. We evaluated the extent to which generated clones and condition screening results were transferable and comparable to results from fully controlled bioreactor systems operated in fed-batch mode at moderate or high cell densities. Direct comparison of 22 different production clones showed great transferability. We observed the same growth and expression characteristics, and identical clone rankings except one host-Fab-leader combination. This outcome demonstrates the explanatory power of HTP µ-bioreactor data and the suitability of this platform as a screening tool in upstream development of microbial systems. Fast, reliable, and transferable screening data significantly reduce experiments in fully controlled bioreactor systems and accelerate process development at lower cost.

High-Throughput Raman Spectroscopy Combined with Innovate Data Analysis Workflow to Enhance Biopharmaceutical Process Development

Raman spectroscopy has the potential to revolutionise many aspects of biopharmaceutical process development. The widespread adoption of this promising technology has been hindered by the high cost associated with individual probes and the challenge of measuring low sample volumes. To address these issues, this paper investigates the potential of an emerging new high-throughput (HT) Raman spectroscopy microscope combined with a novel data analysis workflow to replace off-line analytics for upstream and downstream operations. On the upstream front, the case study involved the at-line monitoring of an HT micro-bioreactor system cultivating two mammalian cell cultures expressing two different therapeutic proteins. The spectra generated were analysed using a partial least squares (PLS) model. This enabled the successful prediction of the glucose, lactate, antibody, and viable cell density concentrations directly from the Raman spectra without reliance on multiple off-line analytical devices and using only a single low-volume sample (50–300 μL). However, upon the subsequent investigation of these models, only the glucose and lactate models appeared to be robust based upon their model coefficients containing the expected Raman vibrational signatures. On the downstream front, the HT Raman device was incorporated into the development of a cation exchange chromatography step for an Fc-fusion protein to compare different elution conditions. PLS models were derived from the spectra and were found to predict accurately monomer purity and concentration. The low molecular weight (LMW) and high molecular weight (HMW) species concentrations were found to be too low to be predicted accurately by the Raman device. However, the method enabled the classification of samples based on protein concentration and monomer purity, allowing a prioritisation and reduction in samples analysed using A280 UV absorbance and high-performance liquid chromatography (HPLC). The flexibility and highly configurable nature of this HT Raman spectroscopy microscope makes it an ideal tool for bioprocess research and development, and is a cost-effective solution based on its ability to support a large range of unit operations in both upstream and downstream process operations.

A Two-Step Strategy that Combines Epigenetic Modification and Biomechanical Cues to Generate Mammalian Pluripotent Cells.

Cell phenotype can be reversed or modified with different methods, with advantages and limitations that are specific for each technique. Here we describe a new strategy that combines the use of chemical epigenetic erasing with mechanosensing-related cues, to generate mammalian pluripotent cells. Two main steps are required. In the first step, adult mature (terminally differentiated) cells are exposed to the epigenetic eraser 5-aza-cytidine to drive them into a pluripotent state. This part of the protocol was developed, based on the increasing understanding of the epigenetic mechanisms controlling cell fate and differentiation, and involves the use of the epigenetic modifier to erase cell differentiated state and then drive into a transient high plasticity window. In the second step, erased cells are encapsulated in polytetrafluoroethylene (PTFE) micro-bioreactors, also known as Liquid Marbles, to promote 3D cell rearrangement to extend and stably maintain the acquired high plasticity. PTFE is a non-reactive hydrophobic synthetic compound and its use permits the creation of a cellular microenvironment, which cannot be achieved in traditional 2D culture systems. This system encourages and boosts the maintenance of pluripotency though bio-mechanosensing-related cues. The technical procedures described here are simple strategies to allow for the induction and maintenance of a high plasticity state in adult somatic cells. The protocol allowed the derivation of high plasticity cells in all mammalian species tested. Since it does not involve the use of gene transfection and is free of viral vectors, it may represent a notable technological advance for translational medicine applications. Furthermore, the micro-bioreactor system provides a notable advancement in stem cell organoid technology by in vitro re-creating a specific micro-environment that allows for the long-term culture of high plasticity cells, namely as ESCs, iPSCs, epigenetically erased cells and MSCs.

Optimization of the Gibco™ CTS™ LV-MAX™ Lentiviral Production System in Stirred Tank Bioreactors

Background & Aim The use of viral vectors for gene therapy applications is an emerging market that has gained considerable clinical traction in recent years. As the demand for gene therapy solutions has increased, the ability to produce viral vectors at manufacturing scale has become more critical. The Gibco™ CTS™ LV-MAX™ Lentiviral Production System is a cGMP-manufactured, xeno-free, 293F-based suspension platform that produces lentiviral vectors with high infectious titers (>1 x 108 TU/mL). The CTS LV-MAX system comprises GMP-banked 293F Virus Production Cells (VPCs), chemically defined growth and production medium, as well as a high-efficiency transfection reagent, titer boosting supplement and novel production enhancer. Since VPCs lack the large T antigen and have been adapted for high density growth in serum-free suspension culture, the LV-MAX system is inherently more scalable and cost-effective than traditional adherent-based virus production platforms. Here, we describe the scale up of the CTS LV-MAX system into stirred tank bioreactors and provide guidance for large scale production of lentiviral vectors under serum-free, suspension conditions. Methods, Results & Conclusion Design of experiment (DOE) and statistical analyses were used to identify the impact of various factors impacting lentiviral production in the CTS LV-MAX System including pH, stir speed, and timing of various reagent additions in both the ambr® 15 microbioreactor system and 3L Thermo Scientific™ HyPerforma™ stirred tank bioreactors. This multi-platform experimental approach allowed for the rapid development of a robust 2L working volume stirred tank bioreactor process capable of generating lentiviral vectors at levels equivalent to those of traditional small scale shake flasks. Future efforts to scale up production into larger bioreactors will further serve to meet the overwhelming production needs currently facing the gene therapy industry.

Ambr® 15 Microbioreactors for CHO Cell Cultivation.

The ambr 15 has become the industry's standard automated microbioreactor system for mammalian cell culture. It has applications throughout the industry, most commonly for cell line screening and media/feed development. On each ambr 15 workstation, conditions in up to 48 × 15 mL bioreactors can be individually controlled while a liquid handler enables automated addition and removal of liquids during the process. Integrated cell counting, metabolite analysis and pH offset correction are also possible thereby reducing the operator interactions that are required. Extensive user and software manuals are supplied by the manufacturer, but in this chapter we describe additional ways of working that we have implemented in routine cell line screening using the ambr 15.

Evaluation of a Small-Scale Perfusion Mimic for Intensified Processes

Genetic Engineering & Biotechnology NewsVol. 39, No. 12 Bioprocess TechnoteEvaluation of a Small-Scale Perfusion Mimic for Intensified ProcessesSartorius Stedim Biotech describes a microbioreactor system that can enable selection of suitable clones during the early stages of process developmentAlison Rees-ManleyAlison Rees-ManleyAlison Rees-Manley (E-mail Address: alison.rees-manley@sartorius-stedim.com) is ambr 15 product manager at at Sartorius Stedim Biotech. Website: www.sartorius-stedim.com.Search for more papers by this authorPublished Online:3 Dec 2019https://doi.org/10.1089/gen.39.12.14AboutSectionsView articleView Full TextPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail View articleFiguresReferencesRelatedDetails Volume 39Issue 12Dec 2019 Information© 2019 by GEN PublishingTo cite this article:Alison Rees-Manley.Evaluation of a Small-Scale Perfusion Mimic for Intensified Processes.Genetic Engineering & Biotechnology News.Dec 2019.52-54.http://doi.org/10.1089/gen.39.12.14Published in Volume: 39 Issue 12: December 3, 2019PDF download

Implementation of a Fully Automated Microbial Cultivation Platform for Strain and Process Screening

Advances in molecular biotechnology have resulted in the generation of numerous potential production strains. Because every strain can be screened under various process conditions, the number of potential cultivations is multiplied. Exploiting this potential without increasing the associated timelines requires a cultivation platform that offers increased throughput and flexibility to perform various bioprocess screening protocols. Currently, there is no commercially available fully automated cultivation platform that can operate multiple microbial fed-batch processes, including at-line sampling, deep freezer off-line sample storage, and complete data handling. To enable scalable high-throughput early-stage microbial bioprocess development, a commercially available microbioreactor system and a laboratory robot are combined to develop a fully automated cultivation platform. By making numerous modifications, as well as supplementation with custom-built hardware and software, fully automated milliliter-scale microbial fed-batch cultivation, sample handling, and data storage are realized. The initial results of cultivations with two different expression systems and three different process conditions are compared using 5 L scale benchmark cultivations, which provide identical rankings of expression systems and process conditions. Thus, fully automated high-throughput cultivation, including automated centralized data storage to significantly accelerate the identification of the optimal expression systems and process conditions, offers the potential for automated early-stage bioprocess development.