Several bacteria cells have been engineered to produce different components of a biopolymer and catalytic enzymes which will be able to trap air pollutants and to degrade some of them.
Our goal being that our project is used as:
- a consumer product that ordinary people can buy
- in a small enclosed device
- on a collective scale (apartment complexes), city or large metropolitan area, especially in developed countries where air pollution is or will become problematic.
Our Chassis organism is E. coli DH5-alpha, BL21DE3 or BL21DE3pLysS. Gene synthesis will be used for providing the coding sequences for expression of the proteins. Sequences were obtained from databases, therefore not requiring extraction of DNA from the originating bacteria.
We are not releasing any GMOs. We are producing a device that can carry our purified components for its final activity. Thus, safe to be released to users. The filter system will degrade toxic components intop less toxic ones, therefore increasing the safety of the trapped materials. The components of our bio-filter are entirely biodegradable (Spider silk, and enzyme proteins).
Level 2 (moderate risk) :
We conducted all of our experimental research in the same level 2 laboratory in the Education center of Institut Pasteur.
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Students were taught a course on common hazards and exposure risks in the lab, including chemical and biological hazards that are found at the Institut Pasteur research laboratories. The course taught us how to prevent exposure to these hazards and emergency response procedures in case of expo-sure. The course also covered our lab waste handling procedures as well as useful information on methods to ensure the research laboratory is free from common physical hazards. Prior to working in the lab, every student in the team received BSL1 and BSL2 lab safety training from Dr Deshmukh Gopaul following guidelines from the Health and safety department of Institut Pasteur. And once in the lab, we received a 2nd training by the laboratory technician of Institut Pasteur Education Center, providing us with specific rules pertaining to waste management in that facility.
All lab work and experiments were done according to laboratory safety policy of the Institut Pasteur.
Safety
Safe Lab Work: From chemicals to electrical equipment, laboratories present a plethora of safety haz-ards, which is why it is so vital to understand the importance of lab safety. The main safety aspects of our daily routine in the lab are the following:
• Using a disinfectant soap to wash your hands before and after working with microorganisms.
• Personal safety: the experiments in the lab were done with gloves, safety glasses, closed leather shoes and lab coats.
• Sterilizing equipment and materials.
• To handle biological materials, we used open bench, biosafety cabinet, cold room -80°C freezer room and also technological platforms.
• Our plates were parafilm sealed and placed in 4°C cold room and cell stocks and cell free extract were stored at -80°C
• Disinfecting work areas before and after use.
• Non-disinfectant soap will remove surface bacteria and can be used if disinfectant soap is not available. Gloves may be worn as extra protection.
• Not eating or drinking in the lab, nor storing food in areas where microorganisms are stored. There are specific eating areas and refrigerators assigned for this purpose.
• Labeling everything clearly. All cultures, chemicals, disinfectant, and media should be clearly and securely labeled with names and dates. If they are hazardous, we label them with proper warning and hazardous information.
• Storing chemicals according to risk categories, flammables, corrosives in specialized cabinets.
• Autoclaving or disinfecting all waste material.
• Cleaning up accidental spills with care.
• The bench was cleaned with ethanol or Anios detergent following each day.
• Collective safety: Common equipment for safety include, fume hoods, Microbiology safety hood, balance area protection, eye wash stations, showers and fire blankets. The equipment have been clearly identified, and operated during our research in the lab; including a fire drill conducted during the summer.
• All the steps for detecting DNA with the agarose gels requested the use of EB (Ethidium bromide), which required particular precautions of manipulation; because the EB is a mutagenic agent: Under hood, on a bench reserved for the use of this product and all the material having touched this bench was not to leave it before cleaning. Specific waste containers were available for liquids and solids.
• The Hygiene and environmental safety office of Institut Pasteur is responsible for laboratory biosafe-ty. http://webcampus.pasteur.fr
• Legifrance is the organism which regulate that govern biosafety in French research laboratories. https://www.legifrance.gouv.fr
The use of bioengineered proteins will minimize the risk of exposure to GMOs. Treatment of pollution by local or federal authorities rather than by individuals could require public "pollution control" regulation enforcement.
Specific risks
Our project did not include the use of gene drives, animals and anti-microbial resistance factors.
Security
The laws and regulations that govern biosafety in research laboratories at the Institut Pasteur are ISO 15189 and ISO/CEI 17025. ISO 15189 is an international standard published by the International Or-ganization for Standardization that specifies requirements for quality and competence particular to medical laboratories. ISO/CEI 17025, is a standard that specifies the general requirements for the competence of calibration and testing laboratories. This standard regarding any laboratory analysis and testing is the repository used during the accreditation audits.
• The laboratory supervisor must enforce the institutional policies that control access to the laborato-ry.
• Students are required to show their vaccination status, compatible for Institut Pasteur, before being allowed to enter lab and perform experiments.
• Security badges are electronic access controls the entry on the campus and specific lab areas.
• Specific biosafety 1/2 clearance is conferred to the Education center Labs.
• Persons must wash their hands before and after working with potentially hazardous materials and before leaving the laboratory.
• Eating, drinking, smoking, handling contact lenses, applying cosmetics, and storing food for human consumption is not permitted in laboratory areas.
• Mouth pipetting is prohibited, mechanical or electric pipetting devices were used.
• Perform all procedures to minimize the creation of splashes and/or aerosols.
• Individual safety equipment, including gloves, lab coat and safety goggles are required.
• Group safety equipment like fume hoods, laminar flow benches are available.
• Decontaminate work surfaces after completion of work and after any spill or splash of potentially infectious material with appropriate disinfectant.
• Decontaminate all cultures, stocks, and other potentially infectious materials before disposal using an effective method.
