title: Hybrid elementary flux analysis/nonparametric modeling: application for bioprocess control.
abstract: BACKGROUND
  The progress in the "-omic" sciences has allowed a deeper knowledge on many biological systems with industrial interest. This knowledge is still rarely used for advanced bioprocess monitoring and control at the bioreactor level. In this work, a bioprocess control method is presented, which is designed on the basis of the metabolic network of the organism under consideration. The bioprocess dynamics are formulated using hybrid rigorous/data driven systems and its inherent structure is defined by the metabolism elementary modes.
  
  
  RESULTS
  The metabolic network of the system under study is decomposed into elementary modes (EMs), which are the simplest paths able to operate coherently in steady-state. A reduced reaction mechanism in the form of simplified reactions connecting substrates with end-products is obtained. A dynamical hybrid system integrating material balance equations, EMs reactions stoichiometry and kinetics was formulated. EMs kinetics were defined as the product of two terms: a mechanistic/empirical known term and an unknown term that must be identified from data, in a process optimisation perspective. This approach allows the quantification of fluxes carried by individual elementary modes which is of great help to identify dominant pathways as a function of environmental conditions. The methodology was employed to analyse experimental data of recombinant Baby Hamster Kidney (BHK-21A) cultures producing a recombinant fusion glycoprotein. The identified EMs kinetics demonstrated typical glucose and glutamine metabolic responses during cell growth and IgG1-IL2 synthesis. Finally, an online optimisation study was conducted in which the optimal feeding strategies of glucose and glutamine were calculated after re-estimation of model parameters at each sampling time. An improvement in the final product concentration was obtained as a result of this online optimisation.
  
  
  CONCLUSIONS
  The main contribution of this work is a novel bioreactor optimal control method that uses detailed information concerning the metabolism of the underlying biological system. Moreover, the method allows the identification of structural modifications in metabolism over batch time.
authors.affiliation: IBET/ITQB, Apartado 12, P-2781-901 Oeiras, Portugal. ana.teixeira@dq.fct.unl.pt <ana.teixeira@dq.fct.unl.pt>
authors.forename: Ana P
authors.initials: AP
authors.lastname: Teixeira
authors.forename: Carlos
authors.initials: C
authors.lastname: Alves
authors.forename: Paula M
authors.initials: PM
authors.lastname: Alves
authors.forename: Manuel J T
authors.initials: MJ
authors.lastname: Carrondo
authors.forename: Rui
authors.initials: R
authors.lastname: Oliveira
mesh_terms: D000465:Algorithms; D000818:Animals; D016466:CHO Cells; D003198:Computer Simulation; D006224:Cricetinae; D003412:Cricetulus; D008657:Metabolic Clearance Rate; D008954:Models, Biological; D015202:Protein Engineering / Q000379:methods*; D020543:Proteome / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D020543:Proteome; D011994:Recombinant Proteins