BioMarin operates a cGMP (current Good Manufacturing Practices) manufacturing facility located in Novato, California. The facility includes dedicated cGMP process manufacturing, quality control, and research and development laboratory space. The facility is licensed by the U.S. Food and Drug Administration and the European Commission to manufacture Aldurazyme® (laronidase), an enzyme replacement therapy (ERT) for the treatment of mucopolysaccharidosis I (MPS I). Today, it is also being used to produce Naglazyme® (galsulfase), an ERT for the treatment of mucopolysaccharidosis VI (MPS VI).
In addition to the cGMP facility, BioMarin continues to operate process development and pilot scale laboratories to further evaluate investigational product candidates.
Manufacturing of Enzyme Replacement Therapies:
Aldurazyme and Naglazyme are recombinant forms of human enzymes, alpha-L-iduronidase and N-acetylgalactosamine 4-sulfatase, respectively. Both products are manufactured using continuous perfusion cell culture—a system that allows for the continual ‘harvest’ (removal) of the desired enzyme—and utilize genetically modified Chinese hamster ovary (CHO) cell lines. It is the harvested enzyme that ultimately becomes the therapeutic product following extensive purification.
The following is an overview of the manufacturing process developed by the research and development staff at BioMarin. The success of this process is made possible through the dedicated and coordinated efforts of five primary groups: Process Development, Manufacturing Sciences, Manufacturing, Quality Assurance, and Quality Control.
Host Cell Expansion/ Cell Culture Inoculation
Frozen host cells that have been genetically engineered to produce a specific enzyme are aseptically added to a sterile flask of growth medium. As the cells multiply, they are transferred to a larger flask to expand the volume and allow room for additional cell growth. This process is repeated until there is enough volume at a specific cell density to inoculate (introduce the cells into) a production-scale bioreactor.
Stringent Environmental Controls
Cell culture inoculation and production occurs in class 100,000 clean rooms. The air in this room is changed through HEPA filters 45 times per hour and is approximately 40 times cleaner than office air. The final purification steps take place under even higher air classifications (Class 10,000). An environmental monitoring program is in place to ensure microorganism and particulate levels are kept within the specified levels.
Cell Culture Production
Inside the bioreactors, cells continue to grow and multiply. The cells are continuously stirred and air and specific nutrients are added through sterile connections to support cell growth. Dissolved oxygen, temperature, pH and nutrient levels are closely monitored and controlled within precise limits to enhance both cell growth and enzyme production. As the cells mature, they begin to secrete the desired enzyme that will later be vialed and administered to patients. Before this can happen, however, the enzyme must first be harvested and purified—removed from the growth medium and separated from the other by-product proteins and molecules—and then undergo final bulk formulation.
High Efficiency Production
Optimizing each step of the manufacturing process is an essential component in maintaining an economically viable process.
To separate the desired enzyme from other proteins and the growth medium, the cell culture fluid is passed sequentially through multiple chromatography columns that utilize differential modes of selectivity. With each step, more impurities are removed, resulting in higher specific activity of the desired enzyme.
An Ultra Filtration/Diafiltration (UF/DF) process is used to formulate the enzyme into a solution that can be safely injected into humans. During UF/DF, the solution is first concentrated and then exchanged with formulation buffer to bring the product and the excipients (other controlled ingredients that enhance the shelf-life stability and improve the product's functional characteristics) to the correct concentrations. Once this is complete, the product is at the exact concentration and composition that will be administered to patients.
The bulk formulation solution is shipped off-site under validated conditions to a specialized fill/finish vendor where it is vialed, capped and labeled under sterile conditions. The labeled vials are stored in quarantine until examined by BioMarin’s Quality Assurance personnel. The vials are sampled and tested for a number of characteristics to ensure safety, purity, potency, identity, quality, and composition. When test results indicate that the final specifications have been met, Quality Assurance releases the vials for distribution and use in patients.