EAGER: Transcriptome analysis of CHO cells to improve productivity and control protein aggregation

Clemson University, Clemson SC

Investigators

Abstract

1218345 Harcum Chinese hamster ovary cells (CHO cells) are used to manufacture a majority of the therapeutic proteins on the market today. In particular, CHO cells are able to express recombinant glycoproteins, including monoclonal antibodies, which require complex post-translational modifications. Even though CHO cells have become the new "Escherichia coli" workhorse cell line, there are still many engineering challenges that must be addressed. These challenges can be calculated when assessing the cost to consumers of glycoprotein therapeutics ($10's to $1000's of dollars (US) per milligram with typical doses of 10's of milligrams). One major production challenge for glycoproteins is protein aggregation. Glycoproteins are prone to aggregation due to the size of these molecules, incomplete glycosylation, high synthesis rates, and free thiols. Fortunately, bioprocess engineers have discovered interim solutions, such as, media additives that can reduce protein aggregation in CHO cell cultures, but more efficient long-term solutions are necessary to stabilize production and reduce costs. The overall goal of this research project is to develop and enhance the toolbox for better understanding protein aggregation beginning with gene expression. Factors that lead to protein aggregation and conditions that reduce protein aggregation will be investigated by interrogating the transcriptome profiles developed from the transcriptome-sequencing component of this project. The CHO cell transcriptome will be made publicly available and characterized to promote genome-wide research in CHO cells. Transcriptome analysis will be used to identify genes that affect and respond to protein aggregation. This project will provide a strong conceptual route for increasing the availability of therapeutic proteins and glycoproteins to the public. The availability of this additional high quality CHO cell transcriptome data to the public will allow for a commensurate community knowledge base that will enable all scholars the ability to contribute to our understanding of bioreactor process effects on this host cell line. Broader impacts also include the establishment and maintenance of a pipeline of educated, motivated students, particularly those from under-represented minority populations to undertake careers in biotechnology to meet this growing demand in the US.