BioNetwork Bioprocessing Center 1800G North Greene Street Technology Enterprise Center (TEC) Greenville, NC 27834 252.75

1. BioNetwork Bioprocessing Center 1800G North Greene Street Technology Enterprise Center (TEC) Greenville, NC 27834 252.757.3759 252.757.3745 fax www.bionetwork.org

2. Staff: • William T. (Bill) Cooper, Manager • bcooper@email.pittcc.edu • Greg Smith, PhD, Curriculum Coordinator • gsmith@email.pittcc.edu • Vallere Shelton, Administrative Assistant • vshelton@email.pittcc.edu

3. What is the Bioprocessing Center? Pitt Community College has established a Bioprocessing Center in conjunction with the statewide BioNetwork initiative. It is one of six specialized biotechnology centers developed by the North Carolina Community College System. This Center is a resource center created to promote biotechnology workforce development.  The ultimate goal is to facilitate the growth of biotechnology firms in North Carolina and the surrounding region. Start-up funding for these centers was provided by Golden LEAF.

4. Why Biotechnology? • Biotechnology is the fastest growing sector of the economy in the State of North Carolina, a state where traditional jobs, such as textiles and tobacco, are by and large extinct. • Biotechnology provides products made from living cells that are useful to all of us. • Biotechnology is the wave of the future as scientific and technological discoveries are made daily. • Biotechnology jobs pay well.

5. Ranking of U.S. States in the Number of Biotechnology Jobs

6. State Impact:

7. State Impact:

8. Growth in Protein Technology:

9. How does Industrial Systems fit in? Because the nature of products created through biotechnology is so valuable to industrial interests, down-time can cost companies millions of dollars in lost profits. Machinery that keeps this industry running needs to be serviced, calibrated, maintained, and cleaned on a regular, expeditious basis. Technicians trained in the culture of biotechnology are more valuable to a company than those that are not. They understand the needs of the industry and have the specialized training to keep the down-time to a minimum. Additionally, they have the expertise to be able to service the equipment without causing undue contamination to the product and the environment.

10. What is Contamination? • The introduction of an unwanted substance to a particular process or environment, either intentionally or accidentally; that which has been introduced to a controlled environment. • Types of contaminants: • Water • Soil • Air • Radiological

11. What is Contamination? • Sources of contamination: • Water (Pseudomonas Aeruginosa) • Soil (Bacillus Subtilis) • Air (any aerosolized form of contamination) • Radiological (any unwanted electromagnetic radiation)

12. What is Contamination? • Prevention of contamination: • Water (filtration or irradiation) • Soil (sanitization and/or disinfection) • Air (high efficiency particulate air filters- ultrafiltration) • Radiological (shielding or non-use)

13. What is Contamination? Particles are measured in microns. If you were to slice a 1-inch cube into 25,400 equal size pieces, each slice would be 1 micron thick (so thin that you could not see it). Since cleanroom facilities contain invisible particles that can cause defects, we need to have a way to determine their sizes and possible sources.

14. What is Contamination? These particles cannot be seen with the naked eye and yet may cause serious loss to our client and to their ability to manufacture devices without defects. We measure the amount of particles in the air with a device known as a particle counter. This helps us to determine when to perform or restrict certain activities.

15. What is Contamination? To combat the tremendous amount of particulate released by the human body, cleanroom personnel are asked to wear special garments that are restrictive and may be uncomfortable if the working conditions (temperature, humidity) are not carefully controlled.

16. What is Contamination? Typical Pollutant Sizes That Cause Down Time

17. What is Contamination? People are dressed in booties, gloves, helmets, hoods and special facemasks that are not part of a normal experience. You are also asked to move slowly, to clean up dirt you cannot see, and to follow practices and procedures that may not immediately make sense.The approach to cleanrooms is designed in steps so as to ensure that every effort has been made not to introduce contaminants into the cleanroom.

18. What is Contamination? "Humans are walking, breathing, waste pits from a microbiological perspective and there is no indication we are going to evolve into anything cleaner."Human dust combined with sebum will tenaciously adhere to and contaminate surfaces, creating contamination problems that are simultaneously bacterial, chemical and particulate.

19. What is Contamination? There is an additional complicating factor - normal human activity. Humans breathe, sneeze, cough, talk and move around. Body and breath temperatures cause heat turbulence in addition to air turbulence.

20. What is Contamination? People slough off huge numbers of particles a half micron and larger and these are propelled around by air and body movement. Some examples of how many 0.5 micron and larger particles can be released by simple movement are shown below. Vast amounts of contamination need to be controlled in a cleanroom environment.

21. What is Contamination? Activity               0.3µm Particles Released

22. What is Contamination? Humans also release elemental chemicals that can cause contamination: * Spittle (saliva): potassium, chloride, phosphorus, magnesium, and sodium * Dandruff: calcium, chloride, carbon, and nitrogen * Perspiration: sodium, potassium, chloride, sulfur, aluminum, carbon, and nitrogen * Fingerprints: sodium, potassium, chloride, and phosphorus Consequently, a critical component of cleanroom management is strict adherence to protocol. Cosmetics are prohibited because in addition to their gross particle generation, cosmetics release iron, aluminum, silicone, carbon, titanium, magnesium, potassium, sulfur, and calcium.

23. What is Contamination Control? Just as humans are the greatest potential contamination risk, they are also the greatest resource for contamination control. A thorough, comprehensive training program detailing all aspects of cleanroom management will empower the cleanroom operators to control the degree of contamination during the production process.

24. What is Contamination Control? Just as all operators must be thoroughly trained in occupational skills, knowledge of the product, and all aspects of the production process, they must also be trained in the unique practices and behaviors required for working in the cleanroom environment. Testing and certification of cleanroom operators and subsequent observation of operators after training assures that the training has been implemented and is effective. Training is mandatory for all personnel (including Management), regardless of frequency of entry or job classification.

25. What is Contamination Control? Personal health and hygiene begins at home with daily bathing or showering, shaving, brushing of teeth and hair, and application of silicone-free skin moisturizers to reduce skin flakes. Make-up, hair gels, hair sprays, aromatic after-shave lotions or body lotions are not cleanroom-compatible. While at work, all employees must wash hands after eating and/or using the toilet. Cleanroom-compatible hand cream may be applied prior to gowning.

26. What is Contamination Control? As mentioned earlier, any activity by the cleanroom operator generates millions of viable and non-viable particles. Consequently, it is imperative to limit talking and actions in the cleanroom to only those required for the manufacture of the product. Running, horseplay, and other non-professional activities are not permitted.

27. What is Contamination Control? • Other behavioral requirements include, but are not limited to, the following: • Smoking is not allowed inside the manufacturing facility including all cleanroom areas. Smokers release particles for at least one-half hour after smoking one cigarette.

28. What is Contamination Control? • Nothing is allowed inside the cleanroom complex which is not required in the cleanroom manufacturing process. This includes personal items such as jewelry or keys, cosmetics, tobacco or matches in any form, and food or drink in any form. • Hair may not be combed in the cleanroom gowning area.

29. What is Contamination Control? • Only cleanroom compatible ball-point pens are allowed inside the cleanroom for recording data on cleanroom compatible paper and clipboards. • While working in the cleanroom, mannerisms such as scratching or hand-rubbing. Cleanroom personnel may not access the inside of the cleanroom uniform.

30. What is Contamination Control? • The use of facial tissues is prohibited in the cleanroom. If one must use a cleanroom compatible non-linting tissue, it must be used only in the gowning area and disposed appropriately in waste receptacle. • All doors must remain closed when not entering or exiting. Emergency doors may be alarmed with a visual and audible alarm to enforce compliance.

31. What is Contamination Control? An integral piece of the cleanroom management program is cleaning of the cleanroom. It is important for all personnel who may work in a cleanroom to understand the critical aspects of maintaining this clean environment.

32. What is Contamination Control? Below is a partial list of some of the commonly known contaminants that can cause problems in some cleanroom environments. It has been found that many of these contaminants are generated from five basic sources. The facilities, people, tools, fluids and the product being manufactured can all contribute to contamination.

33. What is Contamination Control? • Facilities • Walls, floors and ceilings • Paint, caulk and coatings • Construction material (sheet rock, wood, saw dust etc.) • Air conditioning debris • Room air and vapors • Spills and leaks • Rust

34. What is Contamination Control? • People • Skin flakes and oil • Perspiration • Cosmetics and perfume • Spit or saliva • Coughing or sneezing • Clothing debris (lint, fibers etc.) • Hair • Gum, Cough Drops • Food or drink

35. What is Contamination Control? • Tool Generated • Friction and wear particles • Lubricants and emissions • Vibrations • Brooms, mops and dusters • Paper • Cardboard • Duct tape • Permanent Markers • Non-clean room pens

36. What is Contamination Control? • Fluids • Particulates floating in air • Bacteria, organics and moisture • Floor finishes or coatings • Thinners or solvents • Cleaning chemicals • Plasticizers (outgases) • Deionized water

37. What is Contamination Control? • Product generated • Silicon chips • Quartz flakes • Cleanroom debris • Aluminum particles • Packing material • Aerosols and smoke

38. Monitoring Contamination? Airborne Particulate Cleanliness Classes (by cubic meter)

39. Monitoring Contamination?

40. Monitoring Contamination?

41. Monitoring Contamination?

42. Safety in Controlled Environments: • Cleanroom safety is as important (if not • more) than any other facility having • laboratories. Workers in these critical • environments must be conscientious • about not only their own safety, but the • safety of the people benefiting from the • products produced in the environment.

43. Safety in Controlled Environments: • Everything one learns about safety in a • general safety course is applicable in • cleanrooms and the responses would • be the same. However, in cleanrooms • the product is treated with much more • respect and the way one behaves is • more critical than anywhere else.

44. Safety in Controlled Environments: • Exposure to chemicals and biologics • is a risk in cleanrooms just as it is in • any laboratory setting, but less likely • to occur in controlled environments • because of the likelihood of operations • ceasing when maintenance or • calibration procedures are being • performed.

45. Design Considerations and Operation: The method most easily understood and universally applied is the one suggested by the Federal Standard 209E in which the number of particles equal to or greater than 0.5 microns measured in a cubic foot of air designates the class number. For example, a class 100,000 cleanroom limits the concentration of airborne particles equal to or greater than 0.5 microns to 100,000 particles in a cubic foot of air.

46. Design Considerations and Operation: Cleanrooms have evolved into two major types which are differentiated by their method of ventilation - turbulent airflow and laminar airflow cleanrooms. The general method of ventilation used in turbulent airflow cleanrooms is similar to that found in buildings such as offices, schools, malls, manufacturing plants, auditoriums, shops, etc. The air is supplied by an air conditioning system through diffusers in the ceiling.

47. Design Considerations and Operation: • A cleanroom differs from an ordinary ventilated room in three ways: • increased air supply • the use of high efficiency filters • room pressurization

48. Design Considerations and Operation: Increased air supply: The increased air supply is an important aspect of particle control. A typical turbulent airflow cleanroom would have at least 10 air changes per hour and likely have between 20 and 60. This additional air supply is mainly provided to dilute to an acceptable concentration the contamination produced in the room.

49. Design Considerations and Operation: High efficiency filters: High efficiency filters are used to filter the supply air into a cleanroom to ensure the removal of small particles. The high efficiency filters used in cleanrooms are installed at the point of air discharge into the room.

50. Design Considerations and Operation: Room Pressurization: Room pressurization is mainly provided to ensure that untreated air does not pass from dirtier adjacent areas into the cleanroom. The cleanroom is positively pressurized with respect to these dirtier areas. This is done by extracting less air from the room than is supplied to it.