Optimization of Cultivation Conditions in Spin Tubes for CHO Cells Producing Erythropoietin

摘要

This paper describes the optimization of cultivation factor settings, i.e. the shaking raternand working volume in 50 mL spin tubes for a Chinese hamster ovary (CHO) cell linernexpressing recombinant human erythropoietin, using a response D-optimal surfacernmethod. The main objectives of the research were, firstly, to determine a setting inrnwhich the product titer and product quality attributes in spin tubes are equivalent tornthose in 250 mL shake flasks in a seven day batch and, secondly, to find a setting inrnwhich the product titer is maximal. The model for product titer prediction as a functionrnof shaking rate and working volume in the defined design space was successfullyrnapplied to the optimization of cultivation conditions in spin tubes for the tested cell line.rnSubsequently, validation experiments were carried out simultaneously in spin tubes,rnshake flasks and bench scale bioreactors in order to compare cell culture performancernparameters such as growth, productivity and product quality attributes in the form ofrnisoform profiles and glycan antennarity structures. The results of the experimentsrnshowed that similar cell culture performance and product quality could be achieved inrnspin tubes when compared to shake flasks. Additionally, bioreactor titers could bernreproduced in spin tubes at high shaking rates and low working volumes, but withrndiffering product quality. Cultivation at lower shaking rates in spin tubes and shakernflasks produced a glycoprotein with a product quality slightly comparable to that fromrnbioreactors, but with titers being only two thirds.