Nucleotide Sugar Donor Metabolism Network Modeling And The Application Of The Monoclonal Antibody Production

Monoclonal antibody (mAbs) drugs have a very good sales in the market of current international biological medicine, which has huge commercial value. But the reaction mechanisms are uncertain and the production technologies are immature, which leads to cost a lot in mass production of monoclonal antibodies in vitro. Metabolism network model of nucleotide sugar donor (NSDs) is very important model in the production of monoclonal antibody, because nucleotide sugar donors support the oligosaccharides for glycosylation reaction process. Establishing the accurate dynamic model of metabolic network, which can help us comprehend the reaction mechanism and do further quantitative analysis for cell metabolism. This paper mainly discusses the mechanism model and feature analysis for NSDs metabolic network, and the application problems of mAbs production process are studied.First, for NSDs metabolic network mechanism model reconstruction in computer, in this paper, under the basic principles of metabolic network reconstruction, metabolic network diagram is reconstructed and the stoichiometric equation is derived. According to the different mechanisms of each enzyme,60 kinetic reaction rates are derived, and some reaction rate are modified. The enzyme kinetics mechanism and kinetic parameters can be get from the BRENDA database. Based on the principle of dynamic mass conservation,33 NSDs dynamic mass balance equations are built basing on 60 kinetic reaction rates. The complete dynamic mechanism model of NSDs metabolic network is built, and do the reconstruction in computer.Second, in order to ensure the precision of NSDs metabolic network dynamic model, the kinetic parameters are estimated and optimized in NSDs metabolic network dynamic model. In this paper, the initial concentration of enzymes, the initial concentration of nucleoside sugar donors, and part of kinetic parameters are estimated and optimized by genetic algorithm. Comparing and analyzing dynamic model results of NSDs metabolic network basing on optimization parameters with online public access to the experimental data, Due to less actual experimental data, comparison result, to some extent, illustrates the feasibility and accuracy of the model.Finally, analysis of mAbs production process model and control framework in process of mAbs production is put forward. Through the analysis of mAbs production process, mAbs production can be divided into three parts:cell culture process in vitro, nucleotide sugar donor metabolic processes, and glycosylation process in cell. The cell culture model is constructed, and carries on the parameter optimization and model calibration. The analysis of control variables for NSDs metabolic network, the hexose and enzymes as control variables were analyzed, respectively, and get the initial concentration of the enzymes to the production of four kinds of NSDs has great influence, so enzyme can be used as a control variable in further research. Finally, the basic control framework of mAbs production process is presented, which can indicate the direction for the follow-up work.