Team:Technion-Israel/Safety
Introduction
Safety has always been our top concern, both while working in the lab and during the design of our project.
We have divided our safety procedures into 3 general categories: Safety during lab work, Safety in project design and Safety laws and regulations.
Safe Lab Work
All lab work and experiments were done according to the laboratory safety policies at the Technion institute [1]. All team members participated in a week of laboratory training in synthetic biology and cloning techniques, with emphasis on safety and proper laboratory techniques, given by experienced personnel. Upon entering the lab, we received information about the location of fire alarms, safety showers, eye washing stations, fire extinguishers, and fume hoods. Additionally, we have read the MSDS file for each material we have used and made sure we know how to store, handle it and dispose of it.
Lab safety was divided into 3 main parts [2]:
1. Personal safety: All lab workers wore pants, closed toe shoes, lab coats, and gloves while working in the lab.
2. Keeping the lab environment sterilized: Our workbenches were cleaned and sterilized before and after working. When working with the fume hood everything was sterilized with ethanol. The waste was separated into distinguished bins for biologic waste, chemical waste, and regular waste.
3. Separated lab spaces with different safety levels: Open-air benches were designated for regular lab work such as cloning (plasmid purification, restriction analysis, etc). The fume hood was designated for work with hazardous chemicals.
Hazardous Substances
Ethidium bromide - A known carcinogen, used in agarose gel.
Liquid nitrogen - May cause frostbite or cryogenic burns if not handled properly.
Trichloroacetic acid (TCA) – Dangerous material, used for protein precipitation.
Acrylamide – Carcinogen and skin irritant, used for SDS-PAGE preparation.
Hydrogen peroxide – Aggressive oxidizer, used in low concentration for the honey circuit tests.
HCl – Strong acid, Hydrogen chloride forms corrosive hydrochloric acid on contact with water that causes skin burn.
NaOH - Sodium hydroxide is a highly caustic base and alkali that decomposes proteins and may cause severe chemical burns.
Safety Risks
Our final product will be Bee-Free honey made by bacteria. To avoid any risk to our future clients and environment by exposing them to genetically modified organism, we will make some safety precautions:
1. The Bacillus subtilis strain used will be with no sporulation mechanism. The bacteria will grow and secrete the enzymes inside the BioCastle capsule that separate the bacteria from the nectar and only allow the nutrients and proteins to pass through the pores to the outside solution.
2. The high concentrations of hydrogen peroxide, as well as the low pH of the nectar, will hinder the growth of bacteria in case there is a malfunction in the capsule.
3. To make sure that our product is sterile, our BeeFree honey will go through gamma irradiation sterilization according to the FDA safety regulation.
Figure 1: Our BeeFree quality assurance system
Safety laws and regulations
During our project we used two organisms, Escherichia coli as a chassis for the cloning process and Bacillus subtilis for our Honey circuit and protein secretion. Both bacteria are generally recognized as safe (GRAS) by the FDA and belong in risk group 1 [3][4].
B. subtilis is a bacterium that has a long history of safe use as a common host microorganism for enzyme production in the food industry, as described in the FDA's GRAS affirmation (Federal Register Vol 64 Issue 78 of April 23, 1999). Furthermore, the genetic modifications in our proteins are well characterized and specific, and the incorporated DNA does not encode nor express any known harmful or toxic substances. According to the FDA, substances that are GRAS under the conditions of their intended use, do not require premarket approval by FDA [5].
To conclude, once our product will be applicable, the FDA regulation for GRAS substances will allow us to market the BeeFree honey quickly without any hindrance.
- Technion Safety and Health Unit, www.safety.net.technion.ac.il/
- Laboratory biosafety manual, 3rd edition, World Health Organization, Geneva 2004. https://www.who.int/csr/resources/publications/biosafety/Biosafety7.pdf?ua=1
- Escherichia coli K-12 Derivatives Final Risk Assessment, U.S. Environmental Protection Agency 1997. https://www.epa.gov/sites/production/files/2015-09/documents/fra004.pdf
- Bacillus subtilis Final Risk Assessment, U.S. Environmental Protection Agency 1997. https://www.epa.gov/sites/production/files/2015-09/documents/fra009.pdf
- FDA regulations: https://www.fda.gov/food/food-ingredients-packaging/generally-recognized-safe-gras
