The effect of pluronic and microcarriers on the growth, recombinant protein production and glycosylation of CHO cells

Various culture conditions enable the maximization of recombinant protein production from culture processes. Monitoring the productivity (volumetric and specific) gives us an idea of culture processes that are efficient for production of the protein products. However, there are many variables that need to be monitored in order to achieve proper operational platforms for production and for the assurance that the product formed is indeed similar or enhanced compared to usual culture processes used for the production.;An important variable that was shown to be effective for decreasing aggregation of IFN-beta is the use of low temperature for the culture processes. The use of low temperature (32°C) in combination with microcarriers increased the volumetric product yield 5 fold compared to a conventional suspension 37°C batch bioreactor culture. The recombinant protein obtained from cultures where higher product yields were reached was tested for consistency in product glycosylation compared to that obtained in the suspension cultures.;The use of Pluronic F68 (PF68) in the culture medium in static cultures discourages the attachment of cells onto T-flask surfaces; this is shown to be a concentration dependent effect where increased PF68 present decreases surface attachment. PF68 in stirred cultures promotes the increase in cell protection from shear stresses and proteolytic damage. Furthermore, the use of PF68 in culture processes does not affect the glycosylation compared to cultures not supplemented with it.;A procedure was developed to increase recombinant protein production, particularly that of Interferon-beta (IFN-beta) and tissue plasminogen activator (tPA), that utilized the introduction of microcarriers. Volumetric protein yields were found to increase by up to 2.5 fold and specific productivity by up to 5.5 fold compared to corresponding suspension cultures of the respective proteins.