Proteomic Investigation of Recombinant CHO Cells in High Density Culture

摘要

Bioreactor optimization is a complex task, involving both media and process development. This task is further complicated by variations in recombinant CHO cell lines. The intracellular effects of different media or reactor conditions are largely undetermined. Study of intracellular responses could provide information to improve recombinant cell performance in two ways. First, accumulated data could help predict requirements of specific cell lines, increasing the efficiency of media and process optimization. Also, such investigation may identify targets of cell engineering to increase performance of recombinant lines. A proteomics approach was used to study the protein expression changes in a recombinant IgG-producing CHO cell line throughout a fed-batch culture. Growth, productivity and metabolism were monitored, and protein expression profiles of growth and production phases were analyzed by 2-dimensional gel electrophoresis. Differentially expressed proteins were selected using Phoretix 2D Expression software and identified using in-gel tryptic digest and MALDI-MS. This study provides insight into the intracellular processes affecting growth and productivity of recombinant CHO cell lines.