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Plasmid DNA Viral Vectors Recombinant Protein Cell Therapeutics aseptic.html Quality Assurance & Control
Microbial Products Suites | Cell Processing Suites | Mammalian Products Suites | Aseptic Fill Suite | Quality Control Lab
Cell Bank | Raw Materials Holding & Product Quarantine | Sterile Equipment Prep/Waste Decontamination

Mammalian Products

The Mammalian Products manufacturing suites are designed and equipped for manufacturing viral vectors and other biologics that are produced using mammalian cell culture technology (e.g. antibodies). The Mammalian Products suites are both ISO Class 7 (< 352,000 particles/m3, 0.5 micron particles) cleanroom rated which is equivalent to the former Federal Standard 209E class 10,000. The air supply to each room is single-pass air that is filtered with terminal HEPA filters located in the ceiling. The low air returns in these suites also have HEPA filters for containment purposes. In addition, these production rooms are maintained at a negative pressure relative to the surrounding production areas. Medical grade carbon dioxide, oxygen, and nitrogen are provided for cell culture applications as indicated above. In addition, biosafety cabinets are provided in each room for cell cultures and viral vector processing activities.

Mammalian Products 1 will be used primarily for cell culture and harvest. This production suite contains equipment and utilities that are appropriate for culturing a variety of different adherent and non-adherent mammalian cell lines. Adherent cell lines are initially grown in tissue culture flasks and roller bottles. Seed cultures may be started in the Cell Processing Suites and transferred into Mammalian Products 1 for expansion in roller bottles or a bioreactor. A large capacity (29 cu. ft.) reach-in CO2 incubator (model 1950, Forma Scientific) is used for roller bottle culture. A Wheaton Roller Apparatus (model 1862, Wheaton Science Products) placed within the reach-in incubator provides capacity for up to 20 large roller bottles. The incubator is supplied with carbon dioxide from the WCBF distribution system. In addition, carbon dioxide levels, temperature and critical alarms for both the incubator and roller apparatus are monitored by the Equipment Monitoring System.

Scalable bioreactors are also available for culture of both adherent and non-adherent cells. A CYTO3 bioreactor (Osmonics, Minnetonka, MN) is used for projects that will require culturing adherent cell lines in larger cell culture vessels. The culture vessel for the CYTO3 bioreactor consists of a parallel plate single-use vessel that scales to provide up to 85,000 cm2 of surface area for cell growth. The bioreactor is perfused with cell culture media that is circulated through an oxygenator in a closed loop system. A digital control unit monitors and controls media pH, dissolved oxygen, gas flow rates and media flow rates. Conditions within the bioreactor can be maintained over long periods of time to allow cells to grow to high densities. A major benefit of the CYTO3 bioreactor is the use of sterile, single-use culture vessels. This minimizes the issues associated with cleaning validation and contamination control.

A Wave Bioreactor System 20P (Wave Biotech LLC, Bedminster, NJ) is used for suspension culture of mammalian cells at the 1L to 10L scale. The Wave Bioreactor utilizes specially designed plastic bags as the cell culture vessel. The culture bags are also supplied as single-use, sterile reactors that provide a closed system for cell culture. The system consists of a bag holder that agitates the vessel during cell cultivation. A control system with an aeration pump and CO2/air mix controller provides controlled aeration of the cell culture bag.

Mammalian Products 2 will be used primarily for downstream recovery and purification of viral vectors and proteins. Harvested cell culture fluid will be transferred to this room for capture and purification of the target molecule by chromatography. Final concentration and formulation of the target molecule is typically achieved using ultrafiltration and diafiltration processes. Bulk product produced in this area will be stored in a closed system and will be maintained in the Product Quarantine area until it is ready for final formulation and fill. The bulk product will be dispensed into vials in the Aseptic Fill suite. Equipment that will be used in Mammalian Products 2 includes standard chromatography and tangential flow filtration (TFF) set-ups. Chromatography will be performed using closed, disposable systems, where possible, to minimize the risk of contamination. Chromatography media will be dedicated to single project use. The TFF system is comprised of a UltraTec peristaltic pump (SciLog, Middleton, WI) pump system with single-use TFF cartridges.