Industrial choices for protein production by large-scale cell culture

Chu, Lily; Robinson, David K.

doi:10.1016/s0958-1669(00)00197-x

Cited by 493 publications

(290 citation statements)

References 30 publications

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“…Spin-filters (SF) have been proposed as cell retention devices for animal cell culture several decades ago (Himmelfarb et al 1969) and became a popular device for perfusion cultivations both for small-scale studies and large-scale processes (Castilho and Medronho 2002;Chu and Robinson 2001;Figueredo et al 2005). They can be either placed inside the bioreactor (internal spin-filters) or in an external recirculation loop.…”

Section: Introductionmentioning

confidence: 99%

CFD simulation of an internal spin-filter: evidence of lateral migration and exchange flow through the mesh

et al. 2009

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In the present work Computational Fluid Dynamics (CFD) was used to study the flow field and particle dynamics in an internal spin-filter (SF) bioreactor system. Evidence of a radial exchange flow through the filter mesh was detected, with a magnitude up to 130-fold higher than the perfusion flow, thus significantly contributing to radial drag. The exchange flow magnitude was significantly influenced by the filter rotation rate, but not by the perfusion flow, within the ranges evaluated. Previous reports had only given indirect evidences of this exchange flow phenomenon in spin-filters, but the current simulations were able to quantify and explain it. Flow pattern inside the spinfilter bioreactor resembled a typical Taylor-Couette flow, with vortices being formed in the annular gap and eventually penetrating the internal volume of the filter, thus being the probable reason for the significant exchange flow observed. The simulations also showed that cells become depleted in the vicinity of the mesh due to lateral particle migration. Cell concentration near the filter was approximately 50% of the bulk concentration, explaining why cell separation achieved in SFs is not solely due to size exclusion. The results presented indicate the power of CFD techniques to study and better understand spin-filter systems, aiming at the establishment of effective design, operation and scale-up criteria.

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Section: Introductionmentioning

confidence: 99%

CFD simulation of an internal spin-filter: evidence of lateral migration and exchange flow through the mesh

et al. 2009

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“…Microcarriers are used routinely in animal cell culture for the growth of anchorage-dependent cells and are among the established technological platforms for industrial production (8). Yet, there are only sporadic reports on their use in insect cell culture mainly because of the anchorage-independent nature of most cell lines and the one-shot, lytic nature of the BEVS.…”

Section: Introductionmentioning

confidence: 99%

Microcarrier Culture of Lepidopteran Cell Lines: Implications for Growth and Recombinant Protein Production

Ikonomou¹,

Drugmand²,

Bastin

et al. 2002

Biotechnology Progress

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Several microcarrier systems were screened with Sf‐9 and High‐Five cell lines as to their ability to support cell growth and recombinant (β‐galactosidase) protein production. Growth of both cell lines on compact microcarriers, such as Cytodex‐1 and glass beads, was minimal, as cells detached easily from the microcarrier surface and grew as single cells in the medium. Cell growth was also problematic on Cytopore‐1 and ‐2 porous microcarriers. Cells remained attached for several days inside the microcarrier pores, but no cell division and proliferation were observed. On the contrary, insect cells grew well in the interior of Fibra‐Cel disks mainly as aggregates at points of fiber intersection, reaching final (plateau) densities of about 4 × 106 (Sf‐9) and 2.7 × 106 (High‐Five) cells mL‐1 (8 × 106 and 5.5 × 106 cells per cm2 of projected disk area, respectively). Their growth was described well by the logistic equation, which takes into account possible inhibition effects. β‐Galactosidase (β‐gal) production of Sf‐9 cells on Fibra‐Cel disks (infected at 3.3 × 106 cells mL‐1) was prolonged (192 h), and specific protein production was similar to that of high‐density free cell infection. Cultispher‐S microcarriers were found to be a very efficient system for the growth of High‐Five cells, whereas no growth of Sf‐9 cells took place for the same system. Concentrations of about 9 × 106 cells mL‐1 were reached within 120 h, with cell growth in both microcarriers and aggregates, appearance of cellular bridges between microcarriers and aggregates, and eventual formation of macroaggregates incorporating several microcarriers. Specific protein productions after β‐gal baculovirus infection at increasing cell concentrations were almost constant, thus leading to elevated volumetric protein production: final β‐gal titers of 946, 1728, and 1484 U mL‐1 were obtained for infection densities of 3.4, 7.2, and 8.9 × 106 cells mL‐1, respectively.

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“…Mammalian cells continue to be the organism of choice for producing complex biopharmaceuticals because of their ability to perform the desired post translational modifications that are essential for in vivo efficacy (Chu and Robinson 2001). Among the various classes of biopharmaceuticals, monoclonal antibodies are the dominant segment covering a broad range of medical indications and this trends expected to continue in the future (Kozlowski and Swann 2006;Reichert 2009).…”

Section: Introductionmentioning

confidence: 99%

Computer programs for modeling mammalian cell batch and fed-batch cultures using logistic equations

Goudar

2012

Cytotechnology

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A MATLAB Ò toolbox was developed for applying the logistic modeling approach to mammalian cell batch and fed-batch cultures. The programs in the toolbox encompass sensitivity analyses and simulations of the logistic equations in addition to cell specific rate estimation. The toolbox was first used to generate time courses of the sensitivity equations for characterizing the relationship between the logistic variable and the model parameters. Subsequently, the toolbox was used to describe CHO cell data from batch and fed-batch mammalian cell cultures. Cell density, product, glucose, lactate, glutamine, and ammonia data were analyzed for the batch culture while fedbatch analysis included cell density and product concentration. In all instances, experimental data were well described by the logistic equations and the resulting specific rate profiles were representative of the underlying cell physiology. The 6-variable batch culture data set was also used to compare the logistic specific rates with those from polynomial fitting and discrete derivative methods. The polynomial specific rates grossly misrepresented cell behavior in the initial and final stages of culture while those based on discrete derivatives had high variability due to computational artifacts. The utility of logistic specific rates to guide process development activities was demonstrated using specific protein productivity versus growth rate trajectories for the 3 cultures examined in this study. Overall, the computer programs developed in this study enable rapid and robust analysis of data from mammalian cell batch and fed-batch cultures which can help process development and metabolic flux estimation.

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Industrial choices for protein production by large-scale cell culture

Cited by 493 publications

References 30 publications

CFD simulation of an internal spin-filter: evidence of lateral migration and exchange flow through the mesh

CFD simulation of an internal spin-filter: evidence of lateral migration and exchange flow through the mesh

Microcarrier Culture of Lepidopteran Cell Lines: Implications for Growth and Recombinant Protein Production

Computer programs for modeling mammalian cell batch and fed-batch cultures using logistic equations

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