Year: 2010-11

Company: BioMarin Pharmaceuticals

Liaison(s): Jim Michaels, Yvette Tang

Founded in 1997, BioMarin Pharmaceuticals (Novato, CA) is a biotechnology company specializing in treatments for orphan diseases. With over 900 employees worldwide, the company currently has four products on the market, two of which are protein-based drugs, or biologics. Aldurazyme, a recombinant form of alpha-L-iduronidase, is used as an enzyme replacement therapy for patients diagnosed with mucopolysaccharidosis I (MPS I). Naglazyme, a recombinant form of N-acetylalactosamine-4-sulfatase, is used to treat the lysomal storage disorder MPS VI. A third, N-acetylgalactosamine-6-sulfatase is in Phase 3 clinical trials for patients diagnosed with MPS IVA. BioMarin is currently expanding into another lysosomal storage disease, Pompe disease, with their recent acquisition of Zystor Therapeutics. The focus of the BioMarin Team Masters Project was to develop a new fed-batch mammalian cell culture process. This is an experimental project designed to optimize process conditions to maximize cell growth and, hopefully thereby, the titer of a Phase I/II clinical trial product, with a goal of increasing the titer by two fold. (Titer = product concentration at harvest). To accomplish this, the team has leveraged the expertise at the Amgen Bioprocessing Center at KGI, including our new lactate-adaptation technology. This lactate–adaptation technology has been shown to improve process performance by reducing lactic acid production, minimizing the need for base addition for pH control, keeping osmolality low, and allowing for the use of concentrated media for the support of high density CHO (Chinese hamster ovary) cultures. The team performed a series of experiments in shake flasks and bioreactors, testing a variety of process parameters. Variables such as cell age, medium composition, and process parameters (pH, temperature, and pCO2) were explored. To determine product titer, a spectrophotmetric assay developed at BioMarin was performed. The team successfully adapted the cell line to grow to higher cell densities in a chemically defined media. The lactate adaptation strategy was evaluated for the BioMarin cell line and compared to our other fed-batch technologies. Specific challenges included cell line stability as well as balancing cell growth and specific productivity. Additionally, an exploration of nutrient additives known to increase productivity in CHO was conducted.