• COVID‐19
• SARS‐CoV‐2
• Vero
• measles
• microcarriers
• scale up
• single use bioreactors

• Merck & Co., Inc

• Mixing time
• OSRs
• Oxygen transfer
• STCC
• Specific power consumption

• Bioreactors
• Oxygen/metabolism
• Equipment Design

• Adventitious roots
• airlift bioreactors
• bubble column bioreactors
• cell cultures
• organ cultures
• scale-up process
• specialized metabolites
• stirred tank bioreactors

• Bioreactors
• Plant Roots/metabolism
• Plant Roots/growth & development
• Biotechnology/methods
• Cell Culture Techniques/methods

• drug delivery
• engineered exosome
• exosome
• mesenchymal stem cell
• osteoarthritis

• bioreactor
• cell growth
• cell monitoring
• cell viability
• optical coherence tomography

• Bioreactor
• embryogenesis
• immersion culture
• in vitro propagation
• liquid culture
• organogenesis
• somatic embryo
• temporary immersion culture

• COVID‐19
• biomanufacturing
• bioprocessing 4.0
• biotechnology
• vaccines

• bioengineering
• bioreactor
• cell-based meat
• cellular agriculture
• cultivated meat
• cultured meat
• medium
• microcarrier
• scaffold
• sensor
• tissue engineering

• REALSENSE1: Monitoring of cell culture parameters using sensors for biomass and nutrients/metabolites in media: Lab-on-a-Chip (LOC) approach, Good Food Institute 2018 Competitive Grant Program and REALSENSE2: From Lab-On-A-Chip to Custom Bioreactor: Scale Good Food Institute
• 451-03-9/2021-14/200358 Ministry of Education, Science and Technological Development of the Republic of Serbia
• 451-03-68/2021-14/200134 Ministry of Education, Science and Technological Development of the Republic of Serbia

• Euler–Euler simulation
• XcellerexTM XDR-10
• computational fluid dynamics
• mixing time
• one-way Euler–Lagrange simulation
• oxygen mass transfer coefficient
• single-use bioreactor
• surface vortex formation

• T32 EB014836 NIBIB NIH HHS
• UL1 TR001420 NCATS NIH HHS

• Computational fluid dynamics (CFD)
• Dissolved carbon dioxide
• Mixing time
• RANS model
• Surface aeration

• Cell culture
• Enzyme-based optical biosensor
• Glucose monitoring
• Process monitoring

Over the last several decades, plants have been developed as a platform for the production of useful recombinant proteins due to a number of advantages, including rapid production and scalability, the ability to produce unique glycoforms, and the intrinsic safety of food crops. The expression methods used to produce target proteins are divided into stable and transient systems depending on applications that use whole plants or minimally processed forms. In the early stages of research, stable expression systems were mostly used; however, in recent years, transient expression systems have been preferred. The production of the plant itself, which produces recombinant proteins, is currently divided into two major approaches, open-field cultivation and closed-indoor systems. The latter encompasses such regimes as greenhouses, vertical farming units, cell bioreactors, and hydroponic systems. Various aspects of each system will be discussed in this review, which focuses mainly on practical examples and commercially feasible approaches.

• expression system
• molecular farming
• plant-derived protein
• production system
• recombinant protein

Streptomyces clavuligerus is a gram-positive filamentous bacterium notable for producing clavulanic acid (CA), an inhibitor of β-lactamase enzymes, which confers resistance to bacteria against several antibiotics. Here we present a comparative analysis of the morphological and metabolic response of S. clavuligerus linked to the CA production under low and high shear stress conditions in a 2D rocking-motion single-use bioreactor (CELL-tainer ^®) and stirred tank bioreactor (STR), respectively. The CELL-tainer^® guarantees high turbulence and enhanced volumetric mass transfer at low shear stress, which (in contrast to bubble columns) allows the investigation of the impact of shear stress without oxygen limitation. The results indicate that high shear forces do not compromise the viability of S. clavuligerus cells; even higher specific growth rate, biomass, and specific CA production rate were observed in the STR. Under low shear forces in the CELL-tainer^® the mycelial diameter increased considerably (average diameter 2.27 in CELL-tainer^® vs. 1.44 µm in STR). This suggests that CA production may be affected by a lower surface-to-volume ratio which would lead to lower diffusion and transport of nutrients, oxygen, and product. The present study shows that there is a strong correlation between macromorphology and CA production, which should be an important aspect to consider in industrial production of CA.

• Streptomyces
• antibiotic
• bioreactors
• clavulanic acid
• fed-batch
• morphology
• oxygen transfer
• shear stress
• single-use
• stirred tank

In 2004, the FDA published a guideline to implement process analytical technologies (PAT) in biopharmaceutical processes for process monitoring to gain process understanding and for the control of important process parameters. Viable cell concentration (VCC) is one of the most important key performance indicator (KPI) during mammalian cell cultivation processes. Commonly, this is measured offline. In this work, we demonstrated the comparability and scalability of linear regression models derived from online capacitance measurements. The linear regressions were used to predict the VCC and other familiar offline biomass indicators, like the viable cell volume (VCV) and the wet cell weight (WCW), in two different industrially relevant CHO cell culture processes (Process A and Process B). Therefore, different single-use bioreactor scales (50–2000 L) were used to prove feasibility and scalability of the in-line sensor integration. Coefficient of determinations of 0.79 for Process A and 0.99 for Process B for the WCW were achieved. The VCV was described with high coefficients of determination of 0.96 (Process A) and 0.98 (Process B), respectively. In agreement with other work from the literature, the VCC was only described within the exponential growth phase, but resulting in excellent coefficients of determination of 0.99 (Process A) and 0.96 (Process B), respectively. Monitoring these KPIs online using linear regression models appeared to be scale-independent, enabled deeper process understanding (e.g. here demonstrated in monitoring, the feeding profile) and showed the potential of this method for process control.

• Capacitance
• Impedance
• Mammalian CHO cell culture
• PAT
• Process monitoring and control
• Scale-up

Gene therapy and viral vaccine are becoming attractive therapeutic options for the treatment of different malignant diseases. Viral vector productions are often using static culture vessels and small volume stainless steel bioreactors (SSB). However, the yield of each vessel can be relatively low and multiple vessels often need to be operated simultaneously. This significantly increases labor intensity, production costs, contamination risks, and limits its ability to be scaled up, thus, creating challenges to meet the quantities required once the gene therapy or viral vaccine medicine goes into clinical phases or to market. Single-use bioreactor combining with microcarrier provides a good option for viral vector and vaccine production. The goal of the present studies was to develop the microcarrier bead-to-bead expansion and transfer process for HEK293T cells and Vero cells and scale-up the cultures to 50–200 l single-use bioreactors. Following microcarrier bead-to-bead transfer, the peak cell concentration of HEK293T cells reached 1.5 × 10⁶ cells/ml in XDR-50 bioreactor, whereas Vero cells reached 3.1 × 10⁶ cells/ml and 3.3 × 10⁶ cells/ml in XDR-50 bioreactor and XDR-200 bioreactor, respectively. The average growth rates reached 0.61–0.68/day. The successful microcarrier-based scaleup of these two cell lines in single-use bioreactors demonstrates potential large-scale production capabilities of viral vaccine and vector for current and future vaccines and gene therapy.

• Bead-to-bead transfer
• HEK293T cells
• Microcarriers
• Process scale-up
• Single-use Bioreactor
• Vero cells

• air entrainment
• breaking waves
• cell culture
• disposable bioreactor
• oxygen mass transfer

• Cell banking, Fed-batch, Innovation, Mammalian cell culture, Productivity

• Animals
• Bioreactors
• Biotechnology/trends
• Cell Culture Techniques
• Humans
• Recombinant Proteins/biosynthesis
• Recombinant Proteins/supply & distribution

• automation
• bioreactors
• conifer
• scale up
• somatic embryogenesis

• metabolic engineering

• Euler–Euler and Euler–Lagrange approaches
• computational fluid dynamics
• human adipose tissue-derived stromal/stem cells
• humane telomerase reversed transcriptase immortalized hASCs
• microcarrier
• particle image velocimetry/shadowgraphy measurements
• segregated growth model

• CFD
• Orbitally shaken bioreactors
• baffled OSR
• oxygen transfer rate

• Bioreactor
• cell immobilization
• hairy root culture
• membrane technology
• somatic embryogenesis
• temporary immersion systems

Filamentous fungi including Aspergillus niger are cell factories for the production of organic acids, proteins and bioactive compounds. Traditionally, stirred-tank reactors (STRs) are used to cultivate them under highly reproducible conditions ensuring optimum oxygen uptake and high growth rates. However, agitation via mechanical stirring causes high shear forces, thus affecting fungal physiology and macromorphologies. Two-dimensional rocking-motion wave-mixed bioreactor cultivations could offer a viable alternative to fungal cultivations in STRs, as comparable gas mass transfer is generally achievable while deploying lower friction and shear forces. The aim of this study was thus to investigate for the first time the consequences of wave-mixed cultivations on the growth, macromorphology and product formation of A. niger.

We investigated the impact of hydrodynamic conditions on A. niger cultivated at a 5 L scale in a disposable two-dimensional rocking motion bioreactor (CELL-tainer^®) and a BioFlo STR (New Brunswick^®), respectively. Two different A. niger strains were analysed, which produce heterologously the commercial drug enniatin B. Both strains expressed the esyn1 gene that encodes a non-ribosomal peptide synthetase ESYN under control of the inducible Tet-on system, but differed in their dependence on feeding with the precursors d-2-hydroxyvaleric acid and l-valine. Cultivations of A. niger in the CELL-tainer resulted in the formation of large pellets, which were heterogeneous in size (diameter 300–800 μm) and not observed during STR cultivations. When talcum microparticles were added, it was possible to obtain a reduced pellet size and to control pellet heterogeneity (diameter 50–150 μm). No foam formation was observed under wave-mixed cultivation conditions, which made the addition of antifoam agents needless. Overall, enniatin B titres of about 1.5–2.3 g L⁻¹ were achieved in the CELL-tainer^® system, which is about 30–50% of the titres achieved under STR conditions.

This is the first report studying the potential use of single-use wave-mixed reactor systems for the cultivation of A. niger. Although final enniatin yields are not competitive yet with titres achieved under STR conditions, wave-mixed cultivations open up new avenues for the cultivation of shear-sensitive mutant strains as well as high cell-density cultivations.

• Aspergillus niger
• Cyclodepsipeptide
• Enniatin
• Heterologous gene expression
• Macromorphology
• Morphology
• Single-use wave-mixed bioreactor
• Talcum microparticle
• Tet-on system

• Pluronic®
• Sf-21 cells
• baculovirus expression vector system
• shear stress

• Bioprocess monitoring
• Carbon dioxide
• Monitoring device
• Oxygen
• Shake-flask culture

Human mesenchymal stem cell (hMSC)-based therapies are of increasing interest in the field of regenerative medicine. As economic considerations have shown, allogeneic therapy seems to be the most cost-effective method. Standardized procedures based on instrumented single-use bioreactors have been shown to provide billion of cells with consistent product quality and to be superior to traditional expansions in planar cultivation systems. Furthermore, under consideration of the complex nature and requirements of allogeneic hMSC-therapeutics, a new equipment for downstream processing (DSP) was successfully evaluated. This mini-review summarizes both the current state of the hMSC production process and the challenges which have to be taken into account when efficiently producing hMSCs for the clinical scale. Special emphasis is placed on the upstream processing (USP) and DSP operations which cover expansion, harvesting, detachment, separation, washing and concentration steps, and the regulatory demands.

• Advanced therapeutic medicinal product
• Allogeneic
• Good manufacturing practice
• Human mesenchymal stem cells
• Microcarrier
• Single-use devices

Plant molecular farming depends on a diversity of plant systems for production of useful recombinant proteins. These proteins include protein biopolymers, industrial proteins and enzymes, and therapeutic proteins. Plant production systems include microalgae, cells, hairy roots, moss, and whole plants with both stable and transient expression. Production processes involve a narrowing diversity of bioreactors for cell, hairy root, microalgae, and moss cultivation. For whole plants, both field and automated greenhouse cultivation methods are used with products expressed and produced either in leaves or seeds. Many successful expression systems now exist for a variety of different products with a list of increasingly successful commercialized products. This chapter provides an overview and examples of the current state of plant-based production systems for different types of recombinant proteins.

• molecular farming
• recombinant protein expression
• therapeutic proteins
• transient expression
• bioreactor

•

Production process for highly pure polysialic acid is shown.

Polysialic acid (polySia) is a promising molecule for various medical applications (e.g., treatment of inflammatory neurodegenerative diseases).

In this study a complete production process for human-identical α-(2,8)-linked polySia was developed using a disposable bioreactor for cultivation of Escherichia coli K1 and single-use membrane adsorbers for downstream processing (DSP). The cultivation process was optimized to minimize complex media components and a maturation process after cultivation was established. The maturation led to further product release from the cell surface into the supernatant. Afterwards DSP was established using sodium hydroxide treatment combined with anion exchange membrane adsorbers for endotoxin and DNA depletion.

After downstream processing the final product had neither detectable protein nor DNA contamination. Endotoxin content was below 3 EU mg⁻¹. Investigation of the maximal chain length showed no effect of the harsh sodium hydroxide treatment during DSP on the stability of the polySia. Maximal chain length was ∼98 degree of polymerization.

• Endotoxin
• Escherichia coli K1
• Membrane adsorber
• Polysialic acid
• Single-use

• Bioreactor
• Mammalian cells
• Suspension culture
• Transient transfection
• VLP production

• bioreactor
• cell differentiation
• large-scale
• megakaryocyte
• single-use bag

• Recombinant vaccines
• culture mode
• immunogen
• production platform
• virus-like particles

• Animals
• Biotechnology/methods
• Cell Culture Techniques
• Humans
• Plants/metabolism
• Viral Vaccines/metabolism
• Virion/metabolism

• Bioprocess monitoring
• Disposable bioreactors
• Disposable sensors
• Process analytical technology
• Process control
• Sensor systems
• Single‐use