Continuing Your Education

Jan EudyAll my life I have been challenged to provide documented evidence of my competency. While in school, I passed tests to move to the next level. In the workplace, I earned certifications from nationally recognized certification bodies to move forward in my career. I first learned of the NRCM while working at Creighton Memorial St. Joseph Hospital in the microbiology laboratory. After I became a Registered Microbiologist, I was promoted to teach Medical Technology interns laboratory techniques in parasitology, mycobacteriology, and mycology. My favorite part of my job was attending seminars about case studies of patients diagnosed with infectious diseases. I enjoyed solving the case studies with my knowledge of microbiology.

Over the years, my certifications became my credentials to new opportunities in jobs with different companies. In addition to my certification as a Registered Microbiologist with the National Registry for Certified Microbiologists, I am certified as a Medical Technologist with a Specialist in Microbiology with the American Society of Clinical Pathologists, a Certified Quality Auditor with the American Society for Quality, a Provisional Auditor with the RABQSA, a certified HACCP Trainer with the International HACCP Alliance and ServSafe certified.

Currently, I am the Corporate Quality Assurance Manager at Cintas Corporation. One aspect of my job is to stay abreast and compliant with the industry standards in the industries that have cleanrooms and controlled environments. I learn what is required by the various standards and communicate this information to colleagues at Cintas to create quality and risk management programs that will meet Cintas’ customer specifications.

I use the internet and free webinars to get most of my continuing education credits. Since 1998, I have given presentations at industry conferences and since 2003, I have written articles for Controlled Environments magazine so I am researching information for these presentations and articles and learning new facts every day. I enjoy learning the new information and then disseminating it to my colleagues. Success for me is to surround myself with knowledgeable people that can take this information and become more successful.

Jan Eudy, RM(NRCM), Corporate Quality Assurance Manager, Cintas Corporation; she was certified in 1977.

Copyright © National Registry of Certified Microbiologists. Reprinted from The Loop, 2013, Issue 1.

In Terms of One

Semaj McIverI am a DNA Sequencing Scientist at GENEWIZ. Our laboratory, which is a satellite lab, is responsible for cultivating and purifying customer-supplied bacterial plasmids as well as purifying DNA from PCR reactions. We then optimize the DNA to obtain the best quality sequence for customers. We also prepare bacterial colonies that express desired inserts for further analysis upon request. Because of my experience in a multi-lab project while at the North Carolina Department of Agriculture (NCDA), I am also responsible for most quality control testing and documentation in our satellite lab.

A former colleague of mine, and current Registered Microbiologist, first introduced me to the NRCM and suggested that I become a Registrant. This suggestion came one day after a string of long days of working on a method validation. After some deliberation and a bit of apprehension, I decided to attempt certification to demonstrate my competency. While studying for the exam, I found that I had the chance to expound upon current knowledge in Microbiology, as the exam required information outside my normal scope of activities.

Obtaining certification offers many benefits. For example, shortly after becoming a Registrant, I was appointed to the NRCM exam development committee for the Food Safety and Quality Exam. This honor has given me a deeper sense of community with Food Microbiologists across the United States while challenging me to stay abreast of current topics in terms of the exam. Becoming a Registered Microbiologist has been a great accomplishment for me and I am eager to see how it further burgeons my career.

Semaj McIver, RM(NRCM), DNA Sequencing Scientist, GENEWIZ, Research Triangle Park, NC; he achieved his certification in 2011.

Copyright© National Registry of Certified Microbiologists. Reprinted from The Loop, 2013, Issue 1.

Protein Aggregates and Immunogenicity

I work as a microbiologist within the Quality Control unit for a small biopharmaceutical company that manufacturers several protein‐based parenteral products. One of the benefits of working for a small company is your role usually has a broader scope than someone in a larger organization. In addition to validating the standard microbiological release tests for in‐process microbial enumeration, endotoxin and sterility, I have been heavily involved in validating and performing the USP <788> subvisible particulate test by light obscuration for our parenteral products. In recent years, this chapter has come under much scrutiny in its applicability, or lack thereof, to protein‐based parenterals.

One of the drivers for this scrutiny is the risk of immunogenicity to the patient that can be brought on by protein aggregation. Proteins may aggregate for various reasons during manufacture, filling, packaging, and after release. Because of this risk, the FDA has asked many in industry to detect and quantify aggregated protein in their protein‐based drugs and determine the risk, if any, of immunogenicity to the patient. There has been much communication between industry, the FDA, and academia on this issue with no clear consensus on how it should be addressed.

One reason for this lack of consensus is no two protein drugs are alike. The protein in Drug A may aggregate and/or be immunogenic while the aggregated protein in Drug B may not. Currently, the FDA has asked many biologics manufacturers to test for subvisible particles by light obscuration down to the ≥2 micron size (current USP sizes are ≥10 and ≥25 microns) since it is thought that aggregates can be detected in this 2‐10 micron range. It is known that using light obscuration to detect protein aggregation has many drawbacks. For one, protein aggregates are relatively translucent and will result in undercounting. Also, all particles and not just protein aggregates will be counted in this lower range. There is also a lack of an appropriate protein standard for instrument calibration and system suitability purposes. Using light obscuration also raises other questions. What is to be made of results caused by non‐protein particles? How much aggregation is acceptable? Should aggregation studies be part of clinical trial development? These questions and many others have added much difficulty to the validation and performance of the subvisible particulate test for protein‐based parenterals.

In the years since I first started validating subvisible particle counting by light obscuration for protein‐based products, I’ve realized that I’ve had to add validation steps to determine suitable degas and mixing methods for each protein product. And there may be more. Looking to the future there is much interest in micro‐flow imaging technology that is being used more and more by industry for characterization studies. This technology is rapidly becoming more sophisticated in providing an accurate assessment of protein aggregation and may one day become a part of USP<788>. Although the issue of protein aggregates and immunogenicity is challenging and there are often more questions than answers, it is exciting for me to be involved in the development of methods that provide further measures of the safety of the medicines being manufactured.

Karla Aberle M.S., RM (NRCM), QC Microbiology Analyst III, Sigma‐Tau Pharmasource, Inc., Indianapolis, IN. Ms. Aberle achieved her NRCM certification in 2008.

Copyright© National Registry of Certified Microbiologists. Reprinted from The Loop, 2010, Issue 2.

Sterility Failure…Connecting the Dots

If you have never had to deal with the repercussions of a positive sterility result, it may interest you to know how complicated a simple open-drop-incubate test can become. This complexity originates with one simple question, “How did that organism get into the media?” Simple, right? Either the product was not sterile or the media was contaminated during testing. Picking one of these options is far from simple – it is nearly impossible to prove. Here is where I come in.

Surprisingly (for a microbiologist), I get paid to use my imagination. Imagine, for instance, that there was a tiny, invisible particle sitting on the floor of the cleanroom. This particle happened to host a bacterial cell. The analyst enters the cleanroom to perform the testing for the day, and the airflow suddenly changes, causing the particle to float and settle on some supplies. The analyst starts testing. Open packaging, drop sample into media. Open package, drop sample. Open, drop. The analyst reaches for a new lot of forceps and brushes her sleeve against the particle. As she starts testing again, her sleeve brushes the laminar flow hood surface. A media container slides along the hood, picking up the particle. The analyst removes that media container from the hood and grabs a fresh one. Her gloves are now contaminated. She pulls at the sealed packaging to expose the sterile device, docking the particle on the edge of the packaging. Forceps pinch the device, carefully pull it out of the package, just barely brushing against the edge, and the sample is dropped into the media container. That particle tags along into the media, and the organism slowly starts to grow.

Fourteen days later, I get a phone call saying that I have a new sterility failure to investigate. Can I scientifically and objectively re-construct this story from historical data, test videos, and interviews? The answer: Maybe. Investigating a sterility failure is essentially connecting the dots. These dots might not be numbered and some may be missing, but that is essentially what it is.

A microbe ended up in the media – connect the dots back to where it originated. What organisms have we been recovering lately from the room or analyst? How recently was the room cleaned? How does technique during testing look? Was the packaging adequately disinfected? Were the forceps sterile? Along with all these internally focused questions, confidence in the product’s sterility must also be assessed. How well did you validate the sterilization? Were there issues with sterilization this time? Are the organisms resistant? Have there been any manufacturing changes? Was the packaging an effective sterile barrier? The list of questions can go on and on. The art in all of this is knowing which dots are worth connecting.

Imagine that that particle mentioned earlier grew into a colony of Staphylococcus epidermidis. This is a common skin originating organism. The product was manufactured in an environment with S. epidermidis, but was then radiation sterilized. This organism is not known to be resistant to radiation. Bioburden testing shows that the devices, pre-sterilization, harbor some organisms. However, this has remained consistent over the past few years. Sterility testing since the validation has never shown any positives. Packaging was validated and no issues were noted. By this point, you may be thinking that it was the testing laboratory’s fault. According to most regulations and standards, that conclusion is far from justified. This is where the real work (fun) begins. Cleanroom data, such as differential pressures, microbial monitoring, videos, gowning records, cleaning records, maintenance records, sterilizer records, etc. are scrutinized to find evidence of contamination.

Every investigation I perform is unique. Variables combine and contradict in new, intriguing, and usually frustrating ways to paint a picture of contamination. Imagine that growth was observed in the sample as well as the instrument monitor. Did the instrument contaminate the sample, did the sample contaminate the instrument, or did both come into this situation independently? I liken this to the classic chicken or egg question. Which came first? Hard to think about. Even harder to prove.

Sterility testing (open-drop-incubate) can lead to stimulating debates, unanswerable questions, and the occasional headache, all in the name of science. So how did that organism get into the media? We may never know for sure… But I will do my best to find out!

Kevin R. Buckingham, RM(NRCM), Failure Investigator, Nelson Laboratories, Inc., Salt Lake City, UT; he earned his NRCM certification in 2010.

Copyright© National Registry of Certified Microbiologists. Reprinted from The Loop, 2011, Issue 3.

Product Development and Saving Lives

After graduating from the University of Wisconsin‐Milwaukee Master’s Program in Biology, I was offered a product development job at 3M in the Medical Division. A significant portion of my new job required the application of microbiology skills and knowledge. I joined the Henrici Society for Microbiologists, a local microbiology association, to connect with others in the field and keep up on current trends.

In school, I had purposely graduated with a Biology degree, instead of microbiology, which allowed me to take a broader range of courses and graduate earlier. While in graduate school, I knew that product development was my career path, and upon graduation, I chose product development in the health care field as a way to make an impact.

The division I am currently working in is focused on reducing the number of hospital‐associated infections. This vision of saving lives drives what I do every day. I am constantly looking at ways to improve our current products and develop new ones that are of value to our customers.

My day-to-day tasks have many similarities and differences to those in academic research. I design experiments (ELISA, Enzyme Assays, etc.), run them, analyze the results, and make decisions about what to do next. The main difference in product development is that at the end of the project, there is a final product that is sold to a customer. Oftentimes during development, new interesting tangential discoveries are made. These discoveries, unless directly related to the project, are documented and either handed to someone else to pursue or set aside for later. While I am privileged to work at 3M where every employee is encouraged to take some time for side projects, we still need to keep our priority projects on track. There are often strict deadlines and specific deliverables that are required in product development.

I spend a considerable amount of time at the bench, but I also have responsibilities outside of the laboratory. I attend meetings about project planning, company and project updates, and general business meetings. I am also responsible for knowing the standards, journal publications, patent literature, FDA documents, and competitive activities for my projects.

An important part of my role in product development is staying current in my field. The NRCM exam provided me an opportunity to gain additional knowledge and credibility as a microbiologist inside and outside of 3M. While studying for and taking the NRCM exam, I was exposed to many new standards, procedures, and techniques that I had missed by pursing a biology degree.

The NRCM has helped tremendously in my product development role. While I was studying for the exam, I transferred to a new group within the Infection Prevention Division of 3M. I began to work on our biological indicator line of products used by hospital and industrial customers to validate their sterilization process. With this move, it was necessary for me to learn all of the AAMI and USP standards that relate to sterilization processes, a topic also required by the NRCM. Not only was I able to immediately put into practice my new knowledge on sterilization but I was exposed to additional USP and other standards that have helped me.

I would strongly urge any microbiologist who works in product development to take the NRCM certification exam. The NRCM exam expanded my knowledge in microbiology and also introduced a world of techniques, standards, and procedures that I may not have otherwise learned.

Heather Webb, RM(NRCM), Senior Molecular Biologist, 3M Infection Prevention Division, St. Paul, MN; she was certified by the NRCM in 2008.

Copyright© National Registry of Certified Microbiologists. Reprinted from The Loop, 2009, Issue 3.