title: Evaluation of the OSCAR system for the production of monoclonal antibodies by CHO-K1 cells.
abstract: Biopharmaceutical production of complex recombinant protein therapeutics currently relies on mammalian cells. The development of high-yielding stable cell lines requires processes of transfection, selection and adaptation. With several technologies available, selection has been most frequently based on dihydrofolate reductase or glutamine synthetase systems, which can be very time-consuming. Due to the pressure to reduce development costs and speed up time to market, new technologies are emerging, as the promising OSCAR expression system that could provide more rapid development of high-yielding stable cell lines than the traditional systems. However, further evaluation of its application in a wider range of cell types and media is still necessary. In this study, application of OSCAR for the transfection of a CHO-K1 cell line with a monoclonal antibody was evaluated. OSCAR was reasonably fast and simple, without negative impact on cell growth characteristics. However, minigene selection was critical, with only pDWM128 working for the cell line assessed. Initial relatively high levels of production decreased significantly in the first few weeks of passing, remaining relatively stable although with low yield thereafter. The results suggest that more work is required to develop methodologies and prove that OSCAR has significant value to the bioproduction industry.
authors.affiliation: IBB-Institute for Biotechnology and Bioengineering, Centre of Biological Engineering, University of Minho, Campus de Gualtar 4710-057, Braga, Portugal.
authors.forename: Ana Rita
authors.initials: AR
authors.lastname: Costa
authors.forename: Maria Elisa
authors.initials: ME
authors.lastname: Rodrigues
authors.forename: Mariana
authors.initials: M
authors.lastname: Henriques
authors.forename: David
authors.initials: D
authors.lastname: Melton
authors.forename: Philip
authors.initials: P
authors.lastname: Cunnah
authors.forename: Rosário
authors.initials: R
authors.lastname: Oliveira
authors.forename: Joana
authors.initials: J
authors.lastname: Azeredo
mesh_terms: D000630:Aminopterin / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000378:metabolism; D016466:CHO Cells; D049109:Cell Proliferation; D006224:Cricetinae; D003412:Cricetulus; D005822:Genetic Vectors; D019271:Hypoxanthine / Q000378:metabolism; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D013936:Thymidine / Q000378:metabolism; D013997:Time Factors; D014162:Transfection / Q000379:methods*
chemical_list: D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D019271:Hypoxanthine; D007041:Hypoxanthine Phosphoribosyltransferase; D000630:Aminopterin; D013936:Thymidine