Bioprocess scale-up is a fundamental component of process development in the biotechnology industry. When scaling up mammalian cell culture process, it important to consider factors such as mixing time, oxygen transfer, and carbon dioxide removal. In this study, cell-free studies were performed production scale 5,000-L bioreactors evaluate issues. Using current bioreactor configuration, had lower transfer coefficient, longer removal rate than that was observed bench 5- 20-L bioreactors. The...

Markov chain Monte Carlo (MCMC) method was applied to model kinetics of a fed-batch Chinese hamster ovary cell culture process in 5,000-L bioreactors. The kinetic consists six differential equations, which describe dynamics viable density and concentrations glucose, glutamine, ammonia, lactate, the antibody fusion protein B1 (B1). has 18 parameters, were calculated from data, whereas other 12 estimated training data set that comprised seven runs using MCMC method. confirmed two validation...

Large-scale bioprocessing is key to the successful manufacturing of a biopharmaceutical. However, cell viability and productivity are often lower in scale-up from laboratory production. In this study, we analyzed CHO cells, which showed percent viabilities 5-KL production scale bioreactor compared 20-L bench-top under seemingly identical process parameters. An increase copper concentration media 0.02 µM 0.4 led doubling albeit still at level than scale. Combined metabolomics proteomics...

CHO cells express glycoproteins containing both the N-acetylneuraminic acid (Neu5Ac) and minor amounts of N-glycolylneuraminic (Neu5Gc) forms sialic acid. As Neu5Gc is not expressed in humans can be recognized as a foreign epitope, there potential for immunogenicity issues glycoprotein therapeutics. During process development glycosylated fusion protein by cells, number culture conditions were identified that affected content recombinant glycoprotein. Sodium butyrate (SB), well-known...

Abstract This study reports the effects of varying concentrations copper sulfate on metabolic and gene transcriptional profile a recombinant Chinese hamster ovary (CHO) cell line producing an immunoglobulin G (IgG)‐fusion protein (B0). Addition 50 μM significantly decreased lactate accumulation in cultures while increasing viable density titer. These changes could be seen from day 6 became increasingly evident with culture duration. Reducing concentration to 5 retained all above beneficial...

Significant amounts of soluble product aggregates were observed during low-pH viral inactivation (VI) scale-up for an IgG4 monoclonal antibody (mAb IgG4-N1), while small-scale experiments in the same condition showed negligible aggregation. Poor mixing and exposure to low pH identified as root cause. To gain a mechanistic understanding problem, protein aggregation properties studied by varying critical parameters including pH, hold time concentration. Comprehensive biophysical...

Elevation of lactate, ammonia, osmolality, and carbon dioxide to inhibitory levels was reported have adverse effects on cell growth protein productivity in mammalian culture. Multivariate analysis methods were used investigate the roles these repressing metabolites a fed-batch CHO culture for antibody fusion B1 (B1) production. Principal Factor Analysis methodology applied manufacturing-scale data 112 runs, which identified threshold values four as follows: (1) ammonium above 5.1 mM inhibit...

Control of carbon dioxide within the optimum range is important in mammalian bioprocesses at manufacturing scale order to ensure robust cell growth, high protein yields, and consistent quality attributes. The majority bioprocess development work done laboratory bioreactors, which levels are more easily controlled. Some challenges control can present themselves when culture processes scaled up, because accumulation a common feature due longer gas-residence time large bioreactors. A stripping...

Shake flasks and bench-top bioreactors are widely used for cell culture process development, however, performances significantly differ between them. In order to apply the results received from small-scale cultures production scale, it is important understand mechanisms underlying differences various systems. This study analyzes expression patterns of Chinese hamster ovary (CHO) cells producing IgG-fusion protein B0 cultured in shake 5-L by CHO-specific DNA microarrays. The data show that...

Abstract Depth filtration‐based harvesting is widely used in mAb manufacturing to remove cell and process‐related impurities. However, it has not been studied on control of product‐related impurities, which are very critical for product quality. In this article, we the interactions depth filter with high low molecular weight species (HMWs LMWs) their direct removal from culture. The process parameters (filter, loading, temperature, flux) were evaluated adsorption HMWs LMWs by filters....

Abstract Semisteady state cultures are useful for studying cell physiology and facilitating media development. Two semisteady states with a viable density of 5.5 million cells/mL were obtained in CHO compared fed‐batch mode control. In the first state, culture was maintained at 5 mM glucose 0.5 glutamine. The second condition had threefold higher concentrations both nutrients, which led to 10% increase lactate production, 78% ammonia 30% reduction growth rate. differences between two...

A successful tissue culture initiation step often begins with effective explant sterilization. To improve douglas fir bud initiation, five sterilization treatments (20% bleach, 100% 3 second flaming, 5 and self-extinguishing flaming) were evaluated for their effectiveness on winter spring The 20% bleach resulted in the highest percentage of healthy cultures (>90% buds). Spring buds showed a higher level contamination (36%) than did (1%). Successful was also achieved by but injury...

Significant amounts of soluble product aggregates were observed in the low-pH viral inactivation (VI) operation during an initial scale-up run for immunoglobulin-G 4 (IgG4) monoclonal antibody (mAb IgG4-N1). Being earlier development, a scale-down model did not exist, nor was it practical to use costly Protein A eluate (PAE) testing VI process at scale, thus, computational fluid dynamics (CFD)-based high-molecular weight (HMW) prediction developed troubleshooting and risk mitigation. It...

Among many operating parameters that control cell culture environment, appropriate mixing and aeration are crucial for cells to meet oxygen demand in aerobic microbial mammalian production processes. A model-based manufacturing facility fit approach was applied define agitation air flow rates during process scale-up from laboratory manufacturing, of which computational fluid dynamics (CFD) the core modeling tool. The realizable k -ε turbulent dispersed Eulerian two-fluid model used simulate...