Continuous Determination of Glucose and Lactate in a Mammalian Cell Culture Fermentation Process

Abstract

This chapter presents a demonstration of continuous determination of glucose and lactate in a mammalian cell-culture fermentation process. The determination of glucose and lactate in mammalian cell-culture fermentation processes is of great importance for the optimization and the control of the production of monoclonal antibodies. The development of a flow-injection system for the simultaneous on-line determination of glucose and lactate based on glucose dehydrogenase and lactate dehydrogenase covalently immobilized on controlled porous glass and the spectrophotometric or fluorimetric detection of the enzymatically produced NADH is described in the chapter. The samples, discontinuously taken from the harvest stream through a sterile filter, were directly injected into the flow system without any sample pretreatment. Pyruvate, an additional ingredient of fermentation broths interfering with the determination of lactate, is removed up to a concentration of 10 mM by means of a glutamate–pyruvate transaminase column. The whole system is computer-controlled and can be integrated into fermentation control units.