The PYMS GZ China Team understands the importance of safety practices in the lab to ensure the protection of lab members and the environment. In order to ensure proper lab practices, team members took an online course to learn the correct safety procedures.
Prior to conducting our experiments, we became familiar with the lab. During our lab tour, we located and learned how to use fire extinguishers, safety showers, spill kits, and eye-washes. Furthermore, we identified the emergency evacuation route in case of a fire.
We were also taught the proper procedure to dispose of various kinds of waste. Learning the purpose of the different disposal units in the lab, we were taught the autoclaving protocol along with chemical waste protocols specific to the ones we used in our experiment.
Additionally, proper PPE was another important point of safety we diligently followed. Upon entering the wet lab, our members were dressed in the proper PPE: close-toed shoes, long pants that covered the ankles, lab coats, and nitrile gloves. UV resistant lab goggles were donned when we worked with UV light.
To ensure the utmost safety of everyone and everything involved in our project, in addition to the safety course, mentors aided us when working with new equipment or protocols. This would make certain that the correct steps were taken during our experiments.
Our experiment was conducted in a biosafety level 2 lab as we were working with plasmids containing DNA derived from the RBD S-protein in SARS-CoV-2. In addition to the safety measures taken in a biosafety level 1 lab (handwashing, biohazard signs, PPE, mechanical pipetting, daily cleaning.
We are using pseudotyped viruses (Lentivirus) expressing different mutants of the RBD S-protein. This lentivirus is commercially available for purchase.
We are using a human cell line, HEK 293T expressing ACE2. This is also available for purchase.
We are using a non-pathogenic (DH5alpha) strain of E.Coli to genetically engineer and express RBD mutants.
Ingestion of the organisms can cause stomach discomfort. In order to prevent accidental consumption, the lab has designated zones for food and for experiments. To enter the food zone, researchers must wash their hands, take off their lab coat, goggles, and gloves. This will help prevent contamination of the food zone.
The pseudovirus we are constructing is commonly used in scientific laboratory research. It is much less dangerous than normal viruses as virulent components are absent. This allows the pseudovirus to only replicate once. Therefore, infection is possible, but the pseudovirus can be handled safely in a biosafety level 2 lab.