Proteomic profiling of recombinant cells from large-scale mammalian cell culture processes

Abstract

Global expression profiling of mammalian cells used for the production of biopharmaceuticals will allow greater insights into the molecular mechanisms that result in a high producing cellular phenotype. These studies may give insights for genetic intervention to possibly create better host cell lines or even to provide clues to more rational strategies for cell line and process development. In this review I will focus on the contribution of proteomic technologies to a greater understanding of the biology of Chinese hamster ovary cells and other producing cell lines such as NS0 mouse cells.

Supplementation of serum free media with HT is not sufficient to restore growth properties of DHFR−/− cells in fed-batch processes – Implications for designing novel CHO-based expression platforms

“…These platforms include optimized vector systems, selection and amplification protocols, sophisticated and partly automated screening procedures as well as media and process platforms tailor-made for the individual needs of the CHO host cell clone. Moreover, transcriptomics (Nissom et al, 2006;Birzele et al, 2010;Kantardjieff et al, 2010), proteomics (Meleady, 2007;Pascoe et al, 2007) and metabolomics approaches (Nolan and Lee, 2011;Chong et al, 2010) have been used to elucidate the regulatory mechanisms within this host cell and genetic modifications might have been introduced to enhance viability, cellular productivity or other beneficial properties (for review, see also Kuystermans et al, 2007;Mohan et al, 2008). Thus, high titers achieved through effec- tive gene amplification as well as the know-how built around them made DHFR-deficient CHO cells very successful production host cells for the biopharmaceutical industry.…”

Proteomic Profiling of Secreted Proteins from CHO Cells Using Surface-Enhanced Laser Desorption Ionization Time-of-Flight Mass Spectrometry

“…Most of the proteomic studies to date have concentrated on profiling cell lysates from high-producing CHO and other producer cell lines such as NS0 to gain an understanding of the biology of these cells to improve productivity (5)(6)(7)(8)(9). Few studies to date, however, have explored the proteome of extracellularly secreted proteins from mammalian cell lines.…”

Enhanced production of recombinant proteins by a small molecule protein synthesis enhancer in combination with an antioxidant in recombinant Chinese hamster ovary cells

“…Temperature shifting resulting in cell cycle arrest can improve transcription, translation, and cytoskeletal rearrangement, with reduced metabolic activity. The reduced metabolism in the cells is reflected in reduced glucose metabolism and oxygen consumption [9,10]. Cell engineering approaches can target the integration of genes for overexpression such as Bcl-2, a regulatory protein involved in apoptosis-mediated cell death [11].…”