Improved yields of the extracellular domain of the epidermal growth factor receptor produced using the baculovirus expression system by medium replacement following infection.
The extracellular domain of the epidermal growth factor receptor (EGFR) was expressed using the baculovirus expression vector system. The maximum level of the EGFR extracellular domain secreted into the medium in Sf-9 (Spodoptera frugiperda or fall army-worm) cell batch culture was approximately 2.5 micrograms ml-1. In order to increase this yield, a process was developed that included the following sequence of steps: batch growth to maximum cell density, infection of the cells with recombinant virus, and replacement of spent medium. By using this process, the specific yield of recombinant protein, which in batch culture drops when infection is carried out at densities greater than 3 x 10(6) cells ml-1, can be maintained at a maximum in cultures infected at densities of 10(7) cells ml-1 or greater. The process, when applied to 3-1 and 11-1 bioreactor cultures, allowed a maximum volumetric yield of triple the maximum value attainable in batch culture. Spent-medium analysis indicates that medium replacement provides certain nutrients that could otherwise be limiting for recombinant protein production.
Tom
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Animal Cell Engineering Group, Biotechnology Research Institute, National Research Council of Canada, Montréal Québec, Canada.
Debanne
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MT
Bédard
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Caron
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Massie
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Kamen
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D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000378:metabolism; D003470:Culture Media; D066246:ErbB Receptors / Q000096:biosynthesis*; D005947:Glucose / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis*; D018411:Spodoptera
D000596:Amino Acids; D003470:Culture Media; D011994:Recombinant Proteins; D066246:ErbB Receptors; D005947:Glucose