Monoclonal antibody production: viability improvement of RC1 hybridoma cell in different types of bioreactor

Abstract

The study was done to improve the viability of the RC1 hybridoma cell in order to produce more amount of monoclonal antibody (mAb). By using the optimized media, the cell had been cultured in two bioreactor systems which were the MiniPerm and Stirred Tank bioreactor (ST bioreactor), and the results were compared to the one obtained by using the T-Flask bioreactor which was used as a standard. The results showed that the ST bioreactor was able to improve the viability of the cell to the value of 91.8% which was a little bit better than the one obtained by the MiniPerm bioreactor (88.6%) and far better than that of achieved by the T-Flask bioreactor (76.4%). This was well correlated with the good growth performance of the cell in the ST bioreactor with the specific growth rate (l) value of 0.0289 h -1 followed by MiniPerm bioreactor with the value of 0.0243 h -1 and then the T-Flask with the value of 0.0151 h -1 . The low value of doubling time (t d ) obtained in the ST bioreactor (24 h) compared to the one obtained in the MiniPerm (29 h) and T-Flask bioreactor (46 h) had also contributed to the higher value of cell viability. As a result a higher concentration of mAb was able to be produced by the ST bioreactor (0.42 g l -1 ) compared to that of the MiniPerm (0.37 g l -1 ) and T-Flask bioreactor (0.23 g l -1 ).

Technological progresses in monoclonal antibody production systems

“…The traditional and most widely used bioreactor type for mAb production is the stainless steel stirred tank bioreactor (STR) ( Figure 3). 9,10,35 This is mainly due to the broad knowledge and experience acquired from microbial fermentation. 10 This results in better characterized and familiar scale-up principles and mechanical designs for sterilization and cleaning, when compared with other bioreactor types.…”

“…10 This knowhow also facilitates the regulation/approval of the process of production, because the regulatory agencies are experienced with products obtained from STR in different modes of operation. 10 Other advantages of STR include the high mass/gas-transfer coefficient values, 35 the high flexibility 35 in terms of applicable working volumes (up to 15,000 L 37 ), and the suitability for different cell types, operation modes, and products, which results in reduction of costs. 10 Furthermore, good culture conditions such as temperature control, culture pH, dissolved oxygen (DO) level, low agitation rate, and aeration with mixed gas and well-controlled conditions are supported and fulfilled by this bioreactor.…”

“…10 Furthermore, good culture conditions such as temperature control, culture pH, dissolved oxygen (DO) level, low agitation rate, and aeration with mixed gas and well-controlled conditions are supported and fulfilled by this bioreactor. 35 This accurate control of culture conditions allows robust mAb production in STR with most commercially available cell lines. However, the productivities obtained could be further improved if the factors causing stress to the cells could be suppressed, such as shear and bubble damage caused by mechanical agitation and gas sparging.…”

“…136,137 Although the ideal growth temperature for most mammalian cells is 37 C (the body temperature), it has been shown that this may not be the best temperature for recombinant protein/mAb production. Indeed, some studies 138,139 have demonstrated that a lower temperature (around [30][31][32][33][34][35] C) may have a positive outcome in terms of mAb specific productivity, while maintaining or improving product quality. 135 However, the increase of the specific productivity does not often results in improved volumetric productivity, because of the negative effects that low temperatures have in the cell growth rate.…”

Assessment of Kinetic and Metabolic Features of Two Hybridomas in Suspension Culture for Production of Two Monoclonal Antibodies for Blood Typing

“…Therefore, the production of MAbs is mainly carried out in stirred tank bioreactors on an industrial and laboratory scale (Wurm, 2004;Ozturk, 2006;Rodrigues et al, 2009) and under strictly controlled and monitored conditions (Jain and Kumar, 2008). The industrial version consists of a stainless steel tank fitted with a mechanical agitator and automatic temperature, pH and oxygen controllers (Chu and Robinson, 2001;Ozturk, 2006;Mel et al, 2008); one of its laboratory versions, known as spinner flask, is much simpler and consists of a cylindrical glass flask with Teflon® stirrer or actuated magnetically. The spinner flask is operated at fairly constant temperature and pH within a CO 2 incubator.…”