Predictive control of hollow-fiber bioreactors for the production of monoclonal antibodies

Dowd, Jason E.; Weber, Ingrid Távora; Rodríguez, Beatriz Álvarez; Piret, James M.; Kwok, K.E.

doi:10.1002/(sici)1097-0290(19990520)63:4<484::aid-bit12>3.0.co;2-5

Cited by 35 publications

(7 citation statements)

References 24 publications

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“…In the third iteration, HPLC analysis revealed no apparent limitations in any of the 17 measured amino acids; however, there were no increases in the final t-PA concentrations among the three cultures in Figure 1. The addition rates for these serial cultures were calculated using adaptive control of glucose concentrations (38). In the following section, a simpler feeding strategy was developed to maintain constant culture conditions.…”

Section: Resultsmentioning

confidence: 99%

Increased t-PA Yields Using Ultrafiltration of an Inhibitory Product from CHO Fed-Batch Culture

2000

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Fed-batch operation for the production of t-PA using Chinese Hamster Ovary (CHO) cells was optimized using serial and parallel experimentation. The feed, an isotonic concentrate, was improved to obtain 2- to 2.5-fold increases in integrated viable cell days versus batch. With a low glucose inoculum train, the viability index was further increased up to 4.5-fold. Hydrolysates were substituted for the amino acid portion of the concentrate with no significant change in fed-batch results. The concentrate addition rate was based on a constant 4 pmol/cell.day glucose uptake rate that maintained a relatively constant glucose concentration (approximately 3 mM). Increased viable cell indices did not lead to concomitant increases in t-PA concentrations compared to batch. The fed-batch concentrate and feeding strategy were shown to be effective in hybridoma culture, where a 4-fold increase in viable cell index yielded a 4-fold increase in antibody concentration. The half-life of t-PA decreased from 43 to 15 days with decreasing cell viability (from 92% to 71%), but this was not sufficient to explain the apparent t-PA threshold. Instead, the CHO results were explained by a reduction in t-PA production at higher extracellular t-PA concentrations that limited the fed-batch maximum at 35 mg/L for the cell line investigated. Analysis of both the total and t-PA mRNA levels revealed no response to increasing extracellular t-PA concentrations upon exogenous additions. Instead, intracellular t-PA levels were increased, revealing a possible secretory pathway limitation. A new reactor configuration was developed using an acoustic filter to retain the cells in the reactor while an ultrafiltration module stripped t-PA from the clarified medium before the permeate was returned to the reactor. By adding this harvesting step, the t-PA fed-batch production was increased over 2-fold, up to a yield of 80 mg/L.

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Section: Resultsmentioning

confidence: 99%

Increased t-PA Yields Using Ultrafiltration of an Inhibitory Product from CHO Fed-Batch Culture

2000

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“…Many strategies of perfusion culture have been reported, including spin filter (5, 6), hollow‐fiber (7 – 9), packed‐bed and fiber‐bed (10 – 15), ultrasonic filter (16 – 19), centrifugal device (20), flat membrane (21), dialysis membrane (22, 23), and fluidized bed (24) for higher productivities by high cell densities (25, 26). However, most conventional perfusion culture systems have some drawbacks, such as unstable operation and difficult sustenance of perfusion rate in high cell density (4).…”

Section: Introductionmentioning

confidence: 99%

Recombinant Antibody Production by Perfusion Cultures of rCHO Cells in a Depth Filter Perfusion System

Lee

Chang

2008

Biotechnol Progress

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Recombinant Chinese hamster ovary cells, producing recombinant antibody against the human platelet, were cultivated in a depth filter perfusion system (DFPS). When perfusion cultures with working volume of 1 L were operated at perfusion rates of 5/d and 6/d, volumetric antibody productivities reached values 28 and 34 times higher than that of batch suspension culture in Erlenmeyer flasks and 43 and 53 times higher than that of batch culture in a controlled stirred tank reactor, respectively. Perfusion cultures in the DFPS showed stable antibody production over the whole culture period of up to 20 days. In the DFPS, inoculated cells in suspension were entrapped in a few hours within the depth filter matrix by medium circulation and retained there until the void space of the filter matrix was saturated by the cultured cells. After cells in the depth filter matrix reached saturation, overgrown viable cells at a perfusion rate of 5/d or 6/d were continuously collected into waste medium at a density of 2-4 x 10(5) cells/mL, which resulted in stable operation at high perfusion rates, maintaining values of process parameters such as glucose/lactate concentration, pH, and dissolved oxygen concentration. Because the DFPS overcomes most drawbacks observed with conventional perfusion systems, it is preferable to be used as a key culture system to produce monoclonal antibody stably for a long culture period.

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“…In Kurkela et al (1993), daily feed rate step changes by an operator were inaccurate and process deviations resulted. Recently, more advanced predictive modeling based on daily glucose analysis by Dowd et al (1999) improved feed rate specification and bioreactor control by up to 7fold compared to operator specification. Alternatively, the process information can be improved by increasing sampling frequency with automated flow injection analysis.…”

Section: Introductionmentioning

confidence: 99%

“…Van der Pol et al (1995) sampled the process at similar high frequencies, but averaged information and manipulated the process at lower frequencies, improving the basis upon which to manipulate the process. Alternatively in Dowd et al (1999), the control performance was designed a priori and, using predictive models so that from daily sampling, up to 8 manipulations of the process flow rate were performed per day.…”

Section: Introductionmentioning

confidence: 99%

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Consistent perfusion culture production requires reliable cell retention and control of feed rates. An on-line cell probe based on capacitance was used to assay viable biomass concentrations. A constant cell specific perfusion rate controlled medium feed rates with a bioreactor cell concentration of approximately 5 x 10(6) cells mL(-1). Perfusion feeding was automatically adjusted based on the cell concentration signal from the on-line biomass sensor. Cell specific perfusion rates were varied over a range of 0.05 to 0.4 nL cell(-1) day(-1). Pseudo-steady-state bioreactor indices (concentrations, cellular rates and yields) were correlated to cell specific perfusion rates investigated to maximize recombinant protein production from a Chinese hamster ovary cell line. The tissue-type plasminogen activator concentration was maximized ( approximately 40 mg L(-1)) at 0.2 nL cell(-1) day(-1). The volumetric protein productivity ( approximately 60 mg L(-1) day(-1) was maximized above 0.3 nL cell(-1) day(-1). The use of cell specific perfusion rates provided a straightforward basis for controlling, modeling and optimizing perfusion cultures.

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Predictive control of hollow-fiber bioreactors for the production of monoclonal antibodies

Cited by 35 publications

References 24 publications

Increased t-PA Yields Using Ultrafiltration of an Inhibitory Product from CHO Fed-Batch Culture

Increased t-PA Yields Using Ultrafiltration of an Inhibitory Product from CHO Fed-Batch Culture

Recombinant Antibody Production by Perfusion Cultures of rCHO Cells in a Depth Filter Perfusion System

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