The Process Development And Scale-up Of Fed-batch Culture Of Mab Based On The Theory Of QBD

As a core technology for producing antibody drugs,large-scale animal cell culture technology has severely hindered the development and application of protein drugs in China due to limitations in the expression level and production scale.For this purpose,The Chinese hamster ovary cell expressing monoclonal antibodies was used to develope the cell culture process.A fed-batch culture process with high product expression and good product quality was obtained,and a scale up and scale down model was also established.The strategy finally achieved stable and reliable production of monoclonal antibody.First,the cell culture medium was screened.The results showed that the cell growth of Dynamis medium was best and the final titer was 1.53 g/L.Based on this result,the yeastolate hydrolysate was able to increase the antibody expression level,and the final titer reached to 2.5 g/L.Then,process validation was performed on a 2 L bioreactor.The result showed that there's no significant difference between 2 L bioreactor and shake flasks,indicating that this fed-batch culture process was able to scale up to a lab scaled bioreactor.Second,the 2 L and 200 L bioreactor were analyzed to establish a scale up and down strategy.The parameters were divided into two categories: non-volume-related parameters and volume-related parameters.The results showed that the cells performances were identical between 2 L and 200 L bioreactors.The statistical method also proved that the 200 L bioreactor and the 2 L bioreactor are equivalent.Further,six key process parameters: temperature,pH,dissolved oxygen,feeding volume,culture time and seeding density in the culture process was conducted using response surface design.The titer and protein qualitywere set as responses.Based on the calculation and simulation methods,the operating parameters of the operating rangewas determined.Finally,the design space was validated on a 200 L bioreactor,the results showed that the product could be consistently and reliably produced by a 200 L production bioreactor.Based on the theory of quality by design(QbD),a fed-batch culture process with high product expression and good product quality was developed,and the design space of the bioreactor was also obtained.In addition,this article was also a good reference for other cell culture process development using QbD theory.