Compositions and Methods for Increasing Recombinant Protein Yields through the Modification of Cellular Properties

Description of Technology:

This technology relates to compositions and methods for improving the growth characteristics of cells engineered to produce biologically active products such as antibodies or glycosylated proteins. Featured is a method that uses gene candidates (e.g., cdkl3, siat7e, or lama4), or their expressed or inhibited products in cell lines, such as Human Embryonic Kidney (including HEK-293), HeLa, or Chinese Hamster Ovary (CHO). The gene expression modulates growth characteristics, such as adhesion properties, of the cell lines thereby increasing recombinant protein yields and reducing product production costs.

Applications:

This technology may be used to improve production of therapeutic and/or diagnostic compounds, including therapeutic proteins or monoclonal antibodies from mammalian cells. Optimization of mammalian cells for use as expression systems in the production of biologically active products is very difficult. For certain applications, anchorage-independent cell lines may be preferred, whereas for other applications, a cell line that adheres to a surface, e.g. is anchorage-dependent, may be preferable. This technology provides a method for identifying a gene whose expression modulates such cellular adhesion characteristics. This method thus leads to an increase in the expression or yield of polypeptides, including therapeutic biologicals, such as antibodies, cytokines, growth factors, enzymes, immunomodulators, thrombolytics, glycosylated proteins, secreted proteins, and DNA sequences encoding such polypeptides and a reduction in the associated costs of such biological products.

Advantages:

This technology offers the ability to improve yields and reduce the cost associated with the production of recombinant protein products through the selection of cell lines having:

• altered growth characteristics
• altered adhesion characteristics
• altered rate of proliferation
• improvement in cell density growth
• improvement in recombinant protein expression level

Market:

Biopharmaceuticals, including recombinant therapeutic proteins and monoclonal antibody-based products used for in vivo medical purposes and nucleic acid based medicinal products now represent approximately one in every four new pharmaceuticals on the market. The market size has been estimated at $33 billion in 2004 and is projected to reach $70 billion by the end of the decade. The list of approved biopharmaceuticals includes recombinant hormones and growth factors, mAB-based products and therapeutic enzymes as well as recombinant vaccines and nucleic acid based products.
Mammalian cells are widely used expression systems for the production of biopharmaceuticals. Human embryo kidney (including HEK-293) and Chinese hamster ovary (CHO) are host cell of choice. The genes identified in this technology (e.g., cdkl3, sia7e, or lama4) can be used to modify these important cell based systems.
This technology is ready for use in drug/vaccine discovery, production and development. The technology provides methods for identification of specific gene targets useful for altering the production properties of either existing cell lines to improve yields or with new cell lines for the production of therapeutic and or diagnostic compounds from mammalian cells.
Companies that are actively seeking production platforms based on mammalian cell lines that offer high efficiency, high throughput systems for protein production or analysis at lower cost and ease of scale-up would be potential licensors of this technology.

Development Status:

Late Stage-Ready for Production

Patent Status:

DHHS Reference No. E-149-2006, a provisional application has been filed.

Licensing Status:

Available for exclusive or non-exclusive licensing

Licensing Contact:

Dr. Peter Soukas, 301-435-4646, soukasp@od.nih.gov

Collaborative Research Opportunity:

The National Institute of Diabetes and Digestive and Kidney Diseases, Biotechnology Core Laboratory, is seeking parties interested in collaborative research projects directed toward the use of this technology with cells for drug and vaccine production and development, including growth optimization, production and product recovery processes. For more information, please contact Dr. Joseph Shiloach, josephs@intra.niddk.nih.gov, or Rochelle S. Blaustein at Rochelle.Blaustein@nih.gov.

Inventors:

Dr. Joseph Shiloach, NIDDK et.al.

Research Focus and Selected Publications for Principal Investigator -
Biotechnology Core Laboratory

The main research interest of Dr. Shiloach's facility is production and purification of biological products. The unit conducts research and process development in growth optimization, production and product recovery processes, involving both native and recombinant prokaryotes and eukaryotes, concentrating on protein production methods and purification, with an emphasis on scale up. In addition, research is conducted on gene expression and metabolism, especially in E.coli and yeast.

Publications:

1. P Jaluria et al. Application of microarrays to identify and characterize genes involved in attachment dependence in HeLa cells. Metab Eng. 2006 Dec 13, Epub ahead of print, doi:10.1016/j.ymben.2006.12.001. [PubMed abs]
2. Phue JN Shiloach J Transcription levels of key metabolic genes are the cause for different glucose utilization pathways in E. coli B (BL21) and E. coli K (JM109). J Biotechnol (109): 21-30, 2004. [Full Text/Abstract]
3. Trinh LB Phue JN Shiloach J Effect of methanol feeding strategies on production and yield of recombinant mouse endostatin from Pichia pastoris. Biotechnol Bioeng (82): 438-44, 2003. [Full Text/Abstract]
4. Bleckwenn NA Bentley WE Shiloach J Exploring vaccinia virus as a tool for large-scale recombinant protein expression. Biotechnol Prog (19): 130-6, 2003. [Full Text/Abstract]
5. Leppla SH Robbins JB Schneerson R Shiloach J Development of an improved vaccine for anthrax. J Clin Invest (110): 141-4, 2002. [Full Text/Abstract]
6. Noronha SB Yeh HJ Spande TF Shiloach J Investigation of the TCA cycle and the glyoxylate shunt in Escherichia coli BL21 and JM109 using (13)C-NMR/MS. Biotechnol Bioeng (68): 316-27, 2000. [Full Text/Abstract]

Prepared April 2007