Biomedical subjects

Konstantin B Konstantinov

Publications and source records attributed to Konstantin B Konstantinov.

3 recordsLinked to original sources

PubMed2007

Decreased pCO(2) accumulation by eliminating bicarbonate addition to high cell-density cultures.

High-density perfusion cultivation of mammalian cells can result in elevated bioreactor CO(2) partial pressure (pCO(2)), a condition that can negatively influence growth, metabolism, productivity, and protein glycosylation. For BHK cells in a perfusion culture at 20 x 10(6) cells/mL, the bioreactor pCO(2) exceeded 225 mm Hg with approximate contributions of 25% from cellular respiration, 35% from medium NaHCO(3), and 40% from NaHCO(3) added for pH control. Recognizing the limitations to the practicality of gas sparging for CO(2) removal in perfusion systems, a strategy based on CO(2) reduction at the source was investigated. The NaHCO(3) in the medium was replaced with a MOPS-Histidine buffer, while Na(2)CO(3) replaced NaHCO(3) for pH control. These changes resulted in 63-70% pCO(2) reductions in multiple 15 L perfusion bioreactors, and were reproducible at the manufacturing-scale. Bioreactor pCO(2) values after these modifications were in the 68-85 mm Hg range, pCO(2) reductions consistent with those theoretically expected. Low bioreactor pCO(2) was accompanied by both 68-123% increased growth rates and 58-92% increased specific productivity. Bioreactor pCO(2) reduction and the resulting positive implications for cell growth and productivity were brought about by process changes that were readily implemented and robust. This philosophy of pCO(2) reduction at the source through medium and base modification should be readily applicable to large-scale fed-batch cultivation of mammalian cells.

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PubMedDate not supplied

Experimental and theoretical analysis of tubular membrane aeration for Mammalian cell bioreactors.

A combination of experimental and theoretical approaches was used to characterize the dynamics of oxygen transfer in a membrane-aerated bioreactor. Pressure profiles along the length of the membrane at varying entrance and exit pressures were determined by actual experimental measurements, unlike most previous studies that have relied solely on theoretical descriptions of the pressure profile in the tubing. The mass transfer coefficient, k(L)a, was also determined under these conditions and was found to be essentially independent of tubing exit pressure. Measurement of the tubing pressure profile coupled with estimation of k(L)a allowed for computation of the oxygen transfer rate (OTR) along the length of the tubing. A mathematical model that incorporated friction pressure loss and losses due to tubing bending was developed to describe the pressure and hence OTR characteristics of membrane-aerated systems. The applicability of the model was verified by testing it on experimentally measured pressure data, and in all cases the model accurately described experimental data. When tubing properties are known, the mathematical model presented in this study allows for a priori estimation of OTR profiles along the length of the tubing. This information is vital for optimal design and scale-up of membrane-aerated bioreactors for mammalian cell culture.

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PubMedDate not supplied

Logistic equations effectively model Mammalian cell batch and fed-batch kinetics by logically constraining the fit.

A four-parameter logistic equation was used to fit batch and fed-batch time profiles of viable cell density in order to estimate net growth rates from the inoculation through the cell death phase. Reduced three-parameter forms were used for nutrient uptake and metabolite/product formation rate calculations. These logistic equations constrained the fits to expected general concentration trends, either increasing followed by decreasing (four-parameter) or monotonic (three-parameter). The applicability of this approach was first verified for Chinese hamster ovary (CHO) cells cultivated in 15-L batch bioreactors. Cell density, metabolite, and nutrient concentrations were monitored over time and used to estimate the logistic parameters by nonlinear least squares. The logistic models fit the experimental data well, supporting the validity of this approach. Further evidence to this effect was obtained by applying the technique to three previously published batch studies for baby hamster kidney (BHK) and hybridoma cells in bioreactors ranging from 100 mL to 300 L. In 27 of the 30 batch data sets examined, the logistic models provided a statistically superior description of the experimental data than polynomial fitting. Two fed-batch experiments with hybridoma and CHO cells in benchtop bioreactors were also examined, and the logistic fits provided good representations of the experimental data in all 25 data sets. From a computational standpoint, this approach was simpler than classical approaches involving Monod-type kinetics. Since the logistic equations were analytically differentiable, specific rates could be readily estimated. Overall, the advantages of the logistic modeling approach should make it an attractive option for effectively estimating specific rates from batch and fed-batch cultures.

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