What a week! All that work making green fluorescent protein (GFP) comes down to this: manual fill and labeling in BTEC’s aseptic processing suite. Sterility is of paramount importance because this is when the product is put in bottles that will go to patients. Contaminated product could lead to patient harm or death. Just look at a fairly recent example at the New England Compounding Center in which 48 people were killed by a fungal meningitis contamination of a steroid.
The basic idea of aseptic processing is that the sterile filtered product needs to be protected from dirty people and dirty environments. Operators are carefully trained in gowning and cleanroom behaviors. Cleanrooms are extremely clean environments as compared to an average laboratory. Ultimately, the product will come out of the biomanufacturing facility completely sterile and ready to use.
After preparatory lectures we entered the gowning area to be trained in proper gowning procedure. Gowning is a choreographed, multi-step process that, when done properly, completely separates the operator from the cleanroom environment. No microbes, shed skin or hair, dust, debris, etc. will ever come in contact with the product under proper gowning and cleanroom conditions.
We worked in a cleanroom that met Class 10,000 (ISO 7) air quality standards and laminar flow hoods that met class 100 (ISO 5) air quality standards. We manually filled bottles with purified GFP, placed stoppers in the bottles, crimped our caps over the top, and did our weight checks and final fill calculations to determine product loss. Our group was within the acceptable range of product loss during fill. We had one double fill that led to marginal product loss. Our final reconciliation of percent product recovery included weight checks, GFP in Cozzoli manual filler assembly, and other places the GFP came in contact with. The calculation showed that we attained nearly 99% recovery. Not bad for our first time in a fill operation!
Working in a cleanroom was a lot more difficult than I thought it would be. Movements must be slow and deliberate. My goggles kept fogging up on me because my mask was not on right. Conditions are cramped in the laminar flow hood, especially when Environmental Monitoring (EM) comes in right in the middle of your work and sets up a particle counter and active air monitor. I kept leaning my head into the hood to get a better look, but that is a big no-no since I would be introducing particulates and other contaminants into the Class 100 environment. Also, cleanrooms are loud places. There are somewhere around 40 changes of air in a Class 10,000 cleanroom per hour, and the laminar hoods have 60 changes of air per hour. That means lots of fan noise from the HVAC system. We were told that some companies need their entire aseptic processing in Class 100 conditions. It must sound like a wind tunnel.
With filling accomplished we left the cleanroom with our bottles of GFP, de-gowned, and headed for the final step – labeling. Labeling was done in a Class 100,000 room. “Labeling is every bit as important as manufacturing,” our instructor told us. If a drug is made perfectly but is labeled improperly it is deemed “misbranded” by the FDA. Misbranding is a prohibited act under FDA regulations and could leave a company subject to fines or steeper penalties like being shut down. The FDA approves most drug labels. Even changes to existing labels must be approved. Therefore, labeling and label reconciliation are carefully documented and checked by quality control, just like every other step in the biomanufacturing process. One thing I learned was that it is very important that the QA representative be independent of manufacturing to limit potential conflicts of interest.
I must say that I got a great deal of satisfaction seeing my finished product on the table once my group completed reconciliation and got the appropriate signatures from the manufacturing manager and QA. But we were not done yet! We had one last quality control step to do. With the day nearly over, we raced back to the lab with several of our bottles to test our product’s pH, conductivity, and concentration.
I had the most incredible time at BTEC this week. I am looking forward to taking everything I have learned here back to the biomanufacturing program at Laney College. We are starting our Associates of Science (A.S.) degree in biomanufacturing classes this Fall. So, if you have enjoyed reading my blog and would like to work towards a career in biomanufacturing, I encourage you to join our program at Laney College or find a program near you on the Bio-Link website (www.bio-link.org).