HEPA CLEAN ROOM and CABINET BEHAVIOR

How to Avoid Microbial Contamination of Your Products Chair – Fran Rabe Presenters – Diane Kadidlo Fran Rabe

DISCLOSURES Fran Rabe ([email protected]) University of Minnesota Molecular and Cellular Therapeutics Facility Manager, Quality Assurance - No financial or conflicts of interest to report

AGENDA Clean room/hood definitions, operations and use Factors impacting contamination Air flow principles Aseptic processing principles and challenges Aseptic processing solutions

CLEAN ROOMS - Definition Federal Standard 209 definition: An enclose area that controls particulate count (viable and non-viable), humidity and temperature. - To meet the definition of a clean room, the extent of air cleanliness, the particulate count, can not exceed the class of cleanliness. Laminar flow hood and Biological Safety Cabinets can be considered mini-clean rooms by definition

PURPOSE and PROBLEMS OF CLEAN ROOMS, LAMINAR OR BIOLOGICAL SAFETY CABINETS Maintain or reduce bioburden! Contributors to bioburden include: n Facility (flooring, walls, etc.) n Equipment generated n Supply generated n Product generated n People generated

CLEAN ROOM /CABINET PRINCIPLES OF AIR FLOW Laminar = Air molecules moving in parallel paths (vertical flow) Horizontal = Air flows from bank of HEPA filters on wall. Movement towards opposite wall air exhaust Vertical = Air flows from ceiling toward floor of room/cabinet. Air exhausts floor or multiple wall exhausts

BIOLOGICAL SAFETY CABINET/ LAMINAR FLOW HOOD/ CLEAN ROOMS Principle Protection of product from contamination from working space Method Highly filtered air (HEPA) air Positive pressure of space Directional air flow

CLEAN ROOM – HORIZONTAL AIR FLOW HEPA filters placed on wall (horizontal) Air flows horizontally across the space Air exits on opposite wall or ceiling {insert diagram}

CLEAN ROOM VERTICAL AIR FLOW (Floor Exhaust) HEPA filters on ceiling Filter air intake through floor

CLEAN ROOM VERTICAL AIR FLOW With Horizontal Exhaust ……………………………………………………………………………………….

HEPA filters on ceiling

. . .

. . .

Air moves across table Exhaust on wall sides

Exhaust on wall sides

Biological Safety Cabinet – Vertical Flow

Laminar Flow Hood – Horizontal Flow

Biological Safety Cabinet/ Laminar Flow Hood/ Clean Room Purpose System

BSC* LFH* CR*

Prevent particle and micro contamination ü

Protect personnel ü

ü ü

* BSC = Biological Safety Cabinet LFH = Laminar Flow Hood

CR = Clean Room

Protect environment (space)

PEOPLE FACTOR Greatest Contributor Body n Hair n Skin flakes n Respiratory aerosols - bacterial - viral Behavior

PEOPLE PARTICLE PRODUCTON Particle size >0.3μm

Per Minute

Motionless (sitting or standing)

100,000

Head, neck, arm, and leg motion

500,000

Head, neck, arm, leg and foot motion

1,000,000

Sitting to standing or reverse

2,5000,000

Walking @ 2.0 mph (very slow)

5,000,000

Walking @ 3.5 mph (casual)

7,500,000

Walking @ 5.0 mph (fast)

10,000,000

ASEPTIC PROCESSING TECHNIQUE Definition: Techniques or processes used to prevent microbial contamination of an area and/or a product or a QC sample

CLEAN ROOM/CABINET ASEPTIC PROCESSING PRINCIPLES The room/cabinet is not a sterile environment Behavior within these environments need to be “controlled” to ensure product integrity is maintained

CLEAN ROOM/CABINET ASEPTIC PROCESSING PRINCIPLES Reduce bioburden of items brought into clean area prior to reaching the clean(est) area Do not contribute to bioburden of product during processing Particles that are released in the space can be flushed out by the air flow within the space

ESTABLISH CLEAN ZONES Zones that ensure materials/product progressively moves from dirty to clean areas assures bioburden is minimized

SEQUENTIAL MOVEMENT from DIRTIEST to CLEANIST

Non Sterile Materials Containers Product Waste

Dirtiest

Cleaner

Applies to Clean Room or Cabinet Work Environment

Cleanest

Establish and Maintain Sterile Field

CLEAN ROOM/CABINET ASEPTIC PROCESSING SOLUTIONS Avoid clutter n Only have necessary equipment/supplies within space Avoid air turbulence n Strategically place large equipment/materials away from air sources n Liberate air flow

REDUCE CLUTTER

Clutter disrupts air flow and allows dirt to find hiding places Clutter prevents thorough cleaning of the space

ASEPTIC PROCESSING SOLUTIONS Reduce overall bioburden prior to placing item in clean environment- clean it Utilize strategic sequential staging of cleanliness Consider downstream impact of human factor Place equipment/materials strategic related to air flow

ASEPTIC PROCESSING CONTROLS Manage waste n Minimize handling during processing n Ensure aerosols are not released n Cover, where possible

HANDLING WASTE Minimize Aerosols

When possible cover your waste before carrying or pouring Handle waste after your critical processing steps have been performed

ASEPTIC PROCESSING SOLUTIONS Movement n In cabinet, don’t reach arms in and out n Once in clean room, stay there, if you can (utilized circulators) Quantity and Rate of Movement n Limit movement to necessary actions n Move slowly

Movement within Your Space

Avoid in-and-out movement of arms from hood Avoid movement in the clean room where you move from room to room

HORIZONTAL AIR FLOW CABINET PROCESSING SOLUTIONS Arrangement of items: - Items should be > 3 inches from most rear portion of cabinet - Items should be > 6 inches from front edge of cabinet - Large should not be placed towards the rear of the cabinet - Items should be arranged relative to cleanliness (you don’t want the dirtiest item placed upstream of your product).

Re-Examine as You Work Re-examine your workspace periodically Items should be > 3 inches from most rear portion of cabinet Avoid air exhaust obstruction

VERTICAL AIR FLOW PROCESSING SOLUTIONS Do not block air intakes on floor of cabinet Minimize equipment that blocks air intake in clean room

VENT OBSTRUCTION

Never place hands, supplies, drapes over air exhaust

LAMINAR HOOD – HORIZONTAL PROCESSING PRINCIPLES Do not interrupt pathways Strategically place equipment/supplies relative to upstream or downstream of each other

WHAT’s WRONG HERE?

Reorganize the equipment to avoid blocking air exhaust with large equipment

MINDFULL OF AIR STREAM

1. Vertical air flow: Hold syringe/hands to side of open tube 2. Minimize exposure of tube to large portion of the syringe

MAINTAIN STERILE ZONES Not all portions of the sterile-gowned individual are considered as sterile Only sterile area of body should be in contact with sterile fields

RED ZONE/GREEN ZONE of ATTIRE Red Zone is “dirty”: Head, Neck, Waist and down, All of Legs, Feet

Green Zone is “clean”: Arms, front and back, Hands Above waist to chest

Maintain Sterile Zones Movement needs to be strategic with consideration of the sterile and non-sterile zones of the body When more than one person is in the space; walk back to back, rather than front to back Avoid leaning against sterile fields Face forward towards sterile field

MINDFULL of HAND POSITION

SUMMARY Minimize initial and contributing bioburden Control your environment through behavior modification Re-examine your work environment Revisit your environment and process periodically - Use impartial review