Safety

Overview

We understand that safety is always the top priority in any biochemical experiment. This principle has been deeply ingrained in every member of BNU-China from the very beginning of their involvement. We place great emphasis on laboratory safety and have established strict laboratory safety management protocols. Thanks to these efforts, all experiments conducted by BNU-China are ensured to be carried out under fully safe conditions.

Risk Identification

Biosafety risks refer to the potential threats to human health, ecosystem balance, or agricultural production caused by biological factors, such as pathogens, pests, and invasive species. Therefore, it is necessary for us to identify and manage potential biosafety risks in our project to ensure the safety of experiment personnel, the laboratory environment, and public health, as well as to improve the safety and reliability of the experiments. This will lay a foundation for the future public and market adoption of our project results.

In the process of experiment design, through discussion, we have analyzed and identified potential biosafety risks in the following three aspects: our chassis organism, the experimental process, and the risks associated with the product reaching the public after future production.

This year, we chose E. coli BL21(DE3) as our chassis organism, which is known for its high efficiency in expressing foreign proteins. This strain is widely used in structural biology, drug development, vaccine production, and basic scientific research, especially in experiments requiring high-yield expression of recombinant proteins, which meets the needs of our project. E. coli BL21(DE3) is generally considered safe and is used in Biosafety Level 1 (BSL-1) laboratory environments, indicating a low potential safety risk to experiment personnel and the laboratory environment. However, our experiment personnel still need to strictly adhere to operational guidelines and safety protocols to avoid gene leakage or contamination.

Of course, there are many safety risks associated with our experimental operations. Specifically, the engineering modification of MSP involves multiple protein separation, purification, and characterization processes, each of which carries safety hazards related to the use of chemical reagents and the operation of large instruments. In the validation experiments, directly using viruses poses a high risk, and we do not have the appropriate laboratory conditions for this. Therefore, we chose to use the PEG10 protein virus-like particle system as a substitute for virus validation.

We have also conducted a series of evaluations regarding the risks associated with the product's marketization. As a drug, improper use of nanodiscs may lead to side effects on the human body. On one hand, if the specificity of the nanodiscs is not high, they may bind to other normal tissues and cells in the body, reducing drug efficacy and affecting normal physiological activities. On the other hand, if the metabolic cycle of the nanodiscs is too long, it will result in a high level of drug concentration in the blood and tissues, potentially causing harm to the tissues. In our project, we conducted pharmacokinetic modeling of the nanodiscs to mitigate this risk. Additionally, the immunogenicity of the drug to different patient groups is another potential risk. The potential risks of nanodiscs as market drugs can also be avoided through preclinical and clinical trials.

Interview on Safety

To better understand the potential experimental risks in our project, determine whether our experimental content and operations meet a series of laboratory safety regulations, and confirm that our experimental reagents and consumables meet laboratory safety standards, we conducted an interview with our laboratory instructor, Ms. Hao Xiaoran.

First, we learned about the types of experiments that our current laboratory conditions can support. According to Ms. Hao, our lab is an open bench laboratory classified as a P1-level lab, which means experiments involving pathogenic microorganisms are not allowed. In other words, bacteria listed in the national human pathogenic microorganisms registry cannot be used in our experiments. If we wish to conduct experiments related to pathogenic microorganisms, we need to change our project and work on something achievable within the lab's capabilities.

Next, Ms. Hao pointed out the potential safety risks in our project this year. The overall risk of this year's project is relatively higher than in previous years, as we need to express virus-like particles of PEG10. From the perspective of experimental safety protection, although this viral protein is non-toxic, we must wear gloves and lab coats as required, and in special cases, we can operate within a biosafety cabinet. We must sterilize any expressed products or the virus itself promptly and handle them only after thorough sterilization.

Finally, Ms. Hao also emphasized our experimental operations and norms. Regarding experimental materials, the chemical reagents we use must strictly comply with national hazardous chemical regulations. For experimental equipment, such as centrifuges, water baths, and autoclaves, we must operate strictly according to the guidelines. Additionally, Ms. Hao stressed the importance of wearing lab coats when entering the laboratory to minimize potential harm to team members during experimental operations.

Laboratory Safety

To enhance the safety management of the iGEM laboratory, our school has established a strict laboratory access system. Each member participating in wet lab experiments must pass an online assessment formulated by the school, participate in a simulation platform for various experimental operations and emergency plans, and sign a laboratory safety responsibility statement.

Our school has formulated and published laboratory management regulations, and the laboratory safety officer supervises and manages laboratory personnel. To improve the safety of the microbiology laboratory, we have equipped the laboratory with protective equipment such as biosafety cabinets and autoclaves.

The Ministry of Education of China has issued the "Safety Regulations for Higher Education Laboratories," which stipulates the safety management of university laboratories. The Laboratory Safety and Equipment Management Office of Beijing Normal University has also released corresponding management measures, which serve as our laboratory's safety management regulations.

Our laboratory guidelines mainly include the following key points:

1. Pay attention to personal protection when entering the laboratory, including wearing lab coats, closed-toe shoes and socks, medical surgical masks, and latex gloves.

2. Laboratory waste should be collected, stored, and disposed of according to school regulations.

3. Regular laboratory safety inspections should be conducted to eliminate potential safety hazards.

4. Laboratory safety information signs should indicate precautions, protective measures, fire safety points, and the laboratory safety officer.

5. Laboratory instruments and equipment should be equipped with user manuals,and hazardous chemicals should have safety information cards.

6. Laboratory personnel should carefully read the usage instructions for laboratory instruments and chemicals.

7. Laboratory personnel should thoroughly understand emergency measures and strictly adhere to the laboratory management regulations.

Chemical Substances Safety

To ensure the safe management of chemicals in the laboratory, we have implemented a range of strict measures to safeguard team members and facilitate the smooth execution of experiments.

We strictly adhere to the laboratory safety management regulations of Beijing Normal University, ensuring that all experimental processes meet safety standards. All experiments involving potentially hazardous chemicals are conducted in dedicated chemical fume hoods to prevent the leakage of harmful gases and accidental exposure. Additionally, team members must wear appropriate personal protective equipment (PPE) such as gloves, goggles, and lab coats when handling chemicals to minimize the risk of direct contact.

Furthermore, we have conducted thorough inspections of the storage of all chemical reagents in the laboratory. All chemical reagents are categorized and stored according to their properties, with hazardous chemicals stored separately from other chemicals in designated, locked cabinets. We ensure that all chemical reagent containers are clearly labeled and accurately identified, and placed within the laboratory’s line of sight to prevent misuse or accidental exposure.

Moreover, we conduct regular safety inspections of the laboratory, including the storage conditions of chemical reagents, the operational status of chemical fume hoods, and the use of personal protective equipment.

Safety Training

Before the commencement of all experiments, the students responsible for experiments in our team have already undergone safety education on the use of equipment such as autoclaves, laminar flow hoods, biosafety cabinets, fume hoods, PCR machines, and centrifuges. They have also completed two semesters of basic chemistry lab courses. All students in our iGEM team have enhanced their awareness of lab safety and strictly adhere to the laboratory safety regulations during experiments. With the help of our lab instructor, Dr. Hao Xiaoran, we conducted additional laboratory safety education jointly with the iGEM team from Beijing No. 5 High School. The instructor taught the relevant regulations for biological and chemical operations. Experiment students are expected to conduct their experimental explorations according to these regulations, with the assistance and supervision of senior team members. After completing the training for all members of the wet lab team, we passed the virtual simulation assessment for emergency response to safety incidents and signed and confirmed the lab safety agreement on-site.

Under this safety training, our experiments proceeded very smoothly. Over the course of several months of experimentation, no laboratory safety incidents occurred.