Safety is always the priority we need to consider, especially when we turn our project into human practices. Our team is fully compliant with the safety and security policies of iGEM. Safety has been carefully and thoroughly guaranteed in the course of our experiment.
Laboratory Safety
Our team has thoroughly considered the safety issue and potential hazards during the experiment process in the laboratory.
Our experiment is performed in ATLATL laboratory. The laboratory has cabinets suitable for the storage of experiment materials, such as Escherichia Coli, Lactobacillus casei, etc. It is also equipped with fire extinguishers, an eyewash station, and other facilities that ensure laboratory safety.
Before the experiment, all members of our team have taken online training courses about bio-safety instructed by Dr. Fan. We have learned the operation of various apparatuses during the experiment, the action that must be taken when an accident occurs, and the standardization of the experiment, obtaining experimental data, and making qualified conclusions. For the particularity of our experiment, we have learned basic knowledge of biochemical and genetic engineering; basic information of DNA, RNA, protein, and DNA plasmid has been instructed. We have also studied the application of genetic engineering, for instance, enzyme digestion, polymerase chain reaction (PCR), etc.
Our experiment is conducted in the BSL-1 laboratory. Biosafety level 1 (BSL-1) applies to laboratory settings in which personnel work with low-risk microbes poses little to no threat of infection on people. For example, a microbe species typical in the BSL-1 laboratory is a non-pathogenic strain of E. coli. BSL-1 labs also require immediate decontamination after spills. Infection materials are also decontaminated before disposal, generally through the use of an autoclave. BSL-1 labs typically consist of research on benches without the use of special contaminant equipment. They are not required to be isolated from surrounding facilities, and they house activities that require only standard microbial practices, such as:
Mechanical pipetting only (no mouth pipetting allowed)
Safe sharps handling
Avoidance of splashes or aerosols
Daily decontamination of all work surfaces when work is complete
Hand washing
Prohibition of food, drink, and smoking materials in a lab setting
Personal protective equipment, such as safety goggles, gloves, and lab coats or gowns
Our laboratory fulfills the aforementioned standards and meets the requirements of general safety in labs of iGEM.
Material Safety
Our team uses three types of bacteria in the experiment: E. coli BL21 (DE3), E. coli Top10, and Lactobacillus casei ATCC 334, all of which are non-pathogenic. However, during the experiment, any substance getting in touch with bacteria will be contaminated and become biohazardous. Therefore, it needs to be disposed of by certain methods. For liquid-contaminated substances, sodium hypochlorite should be added until the liquid lowers to a concentration of 10%. After being kept stationary for 30 minutes, it should be poured away into the tank followed by a large amount of water. As for solid-contaminated substances, they should be placed under high temperature and pressure until all bacteria are killed, which minimizes the risk of biohazardous substances in the laboratory polluting the environment.
All of the enzymes we use are safe, active, and stable, including, BamH1, Sal1, T4 ligase, etc. Other materials, for example, pGEX-6p-1, pUC57-VP1, pUC57-LTB, are uncontaminated and pose few risks to our health. All of the apparatuses, such as Petri dishes, beakers, and mearuring cylinders, are cleaned or sterilized before use.
Application safety
Our project, EV71 oral vaccine, takes the form of a subunit vaccine with adjuvant. The subunit, VP1 in this case, is not a whole microorganism but an immunological part of the capsid of EV71. While the traditional attenuated vaccine may have a risk of reproducing again or reverting back to pathogenic forms in individuals, the subunit vaccine can largely eliminate this possibility. Also, adjuvant, LTB in this case, can strengthen the effect of the subunit in the GI tract.
In addition, our project, the EV71 vaccine, is delivered orally, which also gives it an edge in terms of safety. Oral vaccination can elicit humoral and cellular immune responses at both systemic and mucosal locations, resulting in a longer and stronger effect than that of the traditional injection-based vaccine.
We design to use Lactobacillus casei to be the living carrier of our final vaccine. According to the official paper issued in 2010 by General Office of the Ministry of Health, L. casei is in the bacteria list that can be used in food, namely no harm to humans.
In conclusion, our experiment satisfies the requirements of safety of iGEM, and our project is practicable.
Reference
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Trapotsis, Arthur. “Biosafety Levels 1, 2, 3 & 4: What's The Difference?” Consolidated Sterilizer Systems, 1 Apr. 2020, https://consteril.com/biosafety-levels-difference/.
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Moyle PM, Toth I. Modern subunit vaccines: development, components, and research opportunities. ChemMedChem. 2013 Mar;8(3):360-76. Epub 2013 Jan 11. PMID: 23316023.
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Online: http://www.nhc.gov.cn/sps/s3593/201004/65839d2d57554dd29ae40a52dca92c74.shtml