Continuous Recombinant Protein Production

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A new applications note is available from INTEGRA Biosciences describing a protocol, developed in conjunction with GSI Creos GmbH (Tokyo, Japan), using the CELLine classic bioreactor for continuous recombinant protein production in Baculovirus-infected SF9 cells.

In recent years, Baculovirus expression vectors have become an efficient and popular method for the production of recombinant proteins in insect cell lines. While recombinant proteins expressed in the cytoplasm of an insect host cell can reach concentrations of up to 1 mg / ml, secreted proteins are usually only found at concentrations below 0.01 mg / ml. Obtaining milligram quantities of a secreted protein using a traditional small scale bioreactor or handling large numbers of standard cell culture disposables is consequently time consuming and costly.

The applications study introduces a new protocol, using a novel disposable flask bioreactor, that cultivates high densities of eukaryotic cells, minimises handling time and produces significant cost savings mainly from a 95% reduction of the required serum supplementation. The protocol enables cultivation of SF9 cells in the 20 ml cell compartment of the CELLine classic bioreactor to a density of up to 1 x 108 cells / ml. The resultant culture can be continuously maintained for several weeks and cells harvested every sixth day. Upon transfection of the SF9 cells, with a baculovirus vector, the high cell densities enable production of recombinant protein titres that are typically 50 times higher than that possible with a standard homogeneous cell culture vessel. In addition because the CELLine classic produces concentrated harvesting volumes, laborious concentration of culture supernatants is avoided offering significant savings in the time, labour and material costs of the downstream processing and purification.

Designed for ease-of-use the CELLine classic utilises proprietary membrane technology to separate off the cultivation chamber with an upper semi-permeable membrane through which nutrient can diffuse and a lower one that allows gases to diffuse. Separate ports allow selective access to the upper nutrient supply chamber and the central cultivation chamber. This novel compartmentalised arrangement means that medium can be exchanged without influencing the function or growth of the cells and gases. Consequently, traditional cell growth limitations brought about by a lack of nutrients or the accumulation of metabolic waste can be overcome just as easily as Oxygen deficiency. Such optimised conditions promote significant cell growth and lead to drastically increased cell densities.

Further information on the CELLine classic and a copy of the applications study may be obtained direct from INTEGRA Biosciences.

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