Preparing Human Gene Transfer Products
NYU’s Institutional Biosafety Committee ensures that research involving human gene transfer products follows procedures modeled on the U.S. Occupational Safety and Health Administration’s guidelines on preparing hazardous drugs. These guidelines establish minimum standard operating procedures for the preparation of human gene transfer products requiring biosafety level 2 containment.
The committee may require more stringent procedures or permit less stringent procedures, depending on the risk assessments of each research registration.
A registered pharmacist, trained in the preparation of human gene transfer products, will provide oversight and maintain required records. Individuals will receive appropriate training before they are allowed to prepare human gene transfer products.
Storage of Human Gene Transfer Products
Products will be stored in locked cabinets with biohazard signs prominently displayed on them.
Access to the preparation area will be restricted when human gene transfer products are being prepared. Signs restricting the access of unauthorized personnel will be displayed prominently on all entrances to the work area. The preparation area will have an eyewash and a sink for hand washing. Eating, drinking, smoking, chewing gum, applying cosmetics, and storing food in the preparation area will be prohibited. Procedures for spills and personnel exposures will be posted in the work area.
Biological Safety Cabinets
Human gene transfer products will be prepared in a Class II biological safety cabinet, which has been certified and maintained in accordance with National Sanitation Foundation Standard No. 49. Arrangements for certification and maintenance will be made through NYU’s Environmental Health and Safety office. The biological safety cabinet will be located in an area with minimal air turbulence, in order to minimize leakage to the environment.
Live viral vectors will not be prepared in the same biological safety cabinet as chemotherapy or other drugs. When feasible, the biological safety cabinet will not be used for other purposes during the course of a human gene transfer study. It will be dedicated exclusively to the study.
The interior of the biological safety cabinet will be thoroughly decontaminated before preparation of a human gene transfer product and after such use. The exhaust fan or blower will be left on during cleaning. The interior surface will be wiped with a 10 percent solution of chlorine bleach, followed by 70 percent alcohol. Spray bottles will not be used, due to the risk of spraying the high efficiency particulate arrestance, or HEPA, filter. Staff will refrain from cleaning the space below the work deck unless it is necessary. If cleaning below the work deck is deemed necessary, staff will follow the instructions detailed in cleanup of small spills and spills in biological safety cabinets.
Personal Protective Equipment
Personnel will wear latex or nitrile gloves that are long enough to cover the gown cuff when preparing human gene transfer products. Personnel will wear double gloves if this does not interfere with their technique. Gloves will be changed hourly or will be changed immediately if they are torn, punctured, or contaminated due to a spill. Hands will be washed with soap and water before gloves are put on and after they are removed. Personnel will avoid touching gloved hands to their faces.
A protective disposable gown made of lint-free, low-permeability fabric with a closed front, long sleeves, and elastic or knit closed cuffs will be worn when gene transfer products are being prepared. The cuffs will be tucked under the gloves. If double gloves are worn, the outer glove will be over the gown cuff and the inner glove will be under the gown cuff. When the gown is removed, the inner glove will be removed last. Gowns and gloves will not be worn outside the preparation area.
Human gene transfer products will be prepared in a biological safety cabinet on a shallow tray covered with a disposable, plastic-backed paper liner. The liner will be changed after preparation is completed for the day. Liners also will be changed after any overt spill. Syringes or intravenous (IV) fittings will be of the Luer-lock type. A sharps container will be in, or immediately adjacent to, the biological safety cabinet for disposal of contaminated glass and sharps. Red bags—biohazard waste containers—will be located immediately adjacent to the biological safety cabinet for disposal of all other contaminated materials, including gloves, gowns, and paper liners.
Personnel will use aseptic technique when preparing human gene transfer products. Manipulations will not be performed close to the front edge of a biological safety cabinet. Unsterilized items will be kept downstream from the working area. Entry into and out of the cabinet will be perpendicular to the front. Rapid lateral hand movements will be avoided.
Gloves and gowns will be donned before work is started in the biological safety cabinet. All items necessary for product preparation will be placed within the biological safety cabinet before work is begun. Extraneous items will be kept out of the work area. Vacuum lines and systems will be protected with disinfectant traps and filters.
All syringes and needles used in the course of preparation will be discarded in sharps containers without being crushed, clipped, or capped. If it is necessary to recap contaminated needles, personnel will use a one-handed technique. If it is necessary to remove contaminated needles from syringes or IV lines, personnel will use a method that minimizes the risk of needle stick, such as forceps removal. IV administration sets will be attached and primed within the biological safety cabinet, prior to addition of the human gene transfer product. If priming must occur at the site of administration, the intravenous line will be primed with a fluid that does not contain human gene transfer product, or a back-flow closed system will be used. Extremes of positive and negative pressure in vials will be avoided, such as attempting to withdraw 10 cc of fluid from a 10-cc vial or placing 10 cc of a fluid into an air-filled 10-cc vial.
Packaging Gene Transfer Products for Internal or External Transport
The outside of bags or bottles containing the prepared human gene transfer product will be wiped with alcohol-moistened gauze. Entry ports will be wiped with moist alcohol pads and capped. Prior to transport, bags, bottles, and syringes will be placed in sealed plastic bags or containers designed to minimize breakage risk. The sealed plastic bags or containers containing the human gene transfer product will be placed in a rigid, leak-proof container with sufficient absorbent material to absorb the human gene transfer product in case of a spill. The container will be labeled with a biohazard label. Personnel who package human gene transfer products for external transport will be trained in shipping and receiving biological materials.
All needles and sharps will be placed in puncture-resistant sharps containers immediately after use. All waste possibly contaminated with human gene transfer products will be disposed as regulated medical waste.
Personnel will immediately remove any contaminated clothing and equipment.
Accidental Inoculation or Skin Contact
Personnel will wash the area with soap and running water for at least 5 minutes, taking care not to vigorously disrupt the skin. They also will take care not to squeeze the inoculation site, since this may increase the risk of spread of the human gene transfer product. Personnel will not use bleach or other strong chemicals to clean an exposure site or wound, because harsh chemicals can damage the skin and increase chances of infection through the skin.
Personnel will flush the eyes with copious amounts of running water for 15 minutes.
After initial treatment, personnel will report to Occupational Health Services at One Park Avenue, third floor, or in off hours to the Ronald O. Perelman Center for Emergency Services and complete an employee occupational illness and injury report. Personnel will notify the principal investigator of the incident, so he or she can file an incident report with the Institutional Biosafety Committee.
Personnel will be trained to clean spills, and will immediately clean-up all spills. Personnel will notify the principal investigator of any large spills, so he or she can file an Incident report with the Institutional Biosafety Committee. A large spill is one that cannot be absorbed with one 4"× 4" gauze pad or one paper towel.
Spill Clean-up Supplies
The following spill clean-up supplies will be kept in or near the preparation area:
- a freshly prepared solution containing a 1:10 dilution of household chlorine bleach and a concentration of 0.6 percent sodium hypochlorite
- protective gloves
- low-permeability gowns
- shoe covers
- eye and face protection
- absorbent towels
- gauze pads
- a sharps container
- a red bag waste container
- a small scoop with which to collect glass fragments
If spill is within a biological safety cabinet, the exhaust fan or blower will be left on during cleaning. Personnel will wear double gloves and closed front gowns. Any broken glass fragments will be picked up using forceps or a small scoop and placed in a sharps container. Spilled material will be covered gently with absorbent towels or gauze pads and flooded with an appropriate disinfectant, such as a solution containing a 1:10 dilution of household chlorine bleach, with a final concentration of 0.6 percent sodium hypochlorite. After the absorbent materials are picked up, the area will be wiped clean with a 10 percent solution of chlorine bleach, followed by water, and then 70 percent ethanol.
Contaminated reusable items, such as glassware and scoops, will be decontaminated with a solution of household chlorine bleach at a 1:10 dilution before being removed from the biological safety cabinet. Used clean-up materials and any other contaminated waste will be discarded in red bags.
Within a biological safety cabinet, if the area underneath the main work surface becomes contaminated, first secure, decontaminate, and remove any items stored in the biological safety cabinet. Decontaminate the top of the work deck. Make sure the drain valve is closed and gently pour a 10 percent bleach solution through the grills into the drain pan. Allow 30 minutes of contact time. Position a container as close to the valve as possible, open the valve to drain into the container. If possible, remove the work deck; remaining disinfectant can be removed using absorbent. Flush or wipe down the area with water to remove bleach residue. Flush water through drain to prevent corrosion. Install work deck back into place. Wash hands.
Clean-up of Large Spills Outside Biological Safety Cabinets
A large spill is one that cannot be absorbed with one 4"× 4" gauze pad or one paper towel. After a large spill, personnel will stop work immediately, avoid inhaling airborne materials, place absorbent material over the spill and instruct others to leave the room immediately. The room will be placed off limits for 30 minutes to allow aerosols to settle and the ventilation system to purge the air.
Personnel will then proceed as they would in the cleanup of small spills and spills inside biological safety cabinets.
Personnel will be trained in the hazards of the specific products, and to properly implement these standard operating procedures