Team:TAS Taipei/Safety

SAFETY

General Safety

Our lab at TAS is classified as Biosafety Level 1, which means the lab uses microbes that pose little to no risk to healthy individuals. Hazardous chemicals and solutions are used at a minimal level and in well ventilated areas such as fume hoods in the TAS research lab. For instance, we do not use EtBr for gel electrophoresis. Instead, we use a safer nucleic acid stain called Seeing Safe DNA Dye. Nevertheless, all chemicals and solutions are still treated with all due respect, care, and caution. MSDS are stored on our lab computers and in a folder on the lab technician's desk.

For the vast majority of experiments, we work with a safe and common lab strain, Escherichia coli (E. coli) K-12 DH5α, and follow safety rules set by our lab instructor. We have our own biosafety committee, which consists of three research teachers Dr. Jonathan Hsu, Mr. Jude Clapper, and Dr. Nicholas Ward. They oversee proper work area conditions by checking on disposal of Petri dishes and liquid wastes, sanitation, and teaching proper laboratory techniques. Our guidelines, taken from National Yang Ming Chiao Tung University's Center of Environmental Protection and Safety, cover many safety rules and procedures ranging from lab specific rules to behavior. For example, we prohibit food, open-toed shoes and drinks in the lab. We also have a thorough clean up procedure. We also have a thorough clean up procedure. For example, all bacterial liquid wastes are bleached and used tips are autoclaved before disposal. Other lab wastes are carefully packaged and then sent to a disposal facility to ensure no hazardous materials are mismanaged.

In Taiwan, there are many laws and regulations regarding biosafety in research labs. These can be found in the National Yang-Ming University's Center of Environmental Protection and Safety and Health page linked here. Guidelines can be found here.

Figure 1 - Picture of our laboratory at TAS where our iGEM experiments were conducted.

Figure 2 - Picture of our laboratory at NYCU where our iGEM experiments were conducted during the summer.

Project Specific Safety

E. coli Strains

We mainly used a common, low risk strain of E. coli, K-12 DH5α, as our chassis organism (Environmental Protection Agency, 1977). We genetically engineered the bacteria to produce our desired enzymes. If bacteria were to escape the lab, they would pose low risk to the surrounding community. After use, all bacterial wastes were either autoclaved or bleached to kill the microbes before disposal.

Porcine Blood Testing

We used porcine blood to test our enzyme functionality and provide a model for human red blood cells. Prior to usage, we submitted a check-in form to confirm that we are able to use porcine blood in our lab. After confirmation, we obtained the blood from a farm in Hsinchu, a county in Taiwan, specializing in breeding pigs for scientific research purposes. With a reliable source and robust handling and screening procedures, we made sure our experiments were conducted in a safe manner and that all porcine blood was bleached to kill live cells before disposal.

Coercive Chemicals for Enzyme Functional Tests

While performing enzyme functional tests using Thin Layer Chromatography, we used several strong acids to create solvents and visualizing reagents, including hydrochloric acid, sulfuric acid, and boric acid. Strong acids can cause chemical burns if they come in contact with one’s skin, and can damage the respiratory system if inhaled (Fred Hutch Extranet, n.d.). To ensure safety, we made all solutions and performed tests in the chemical fume hood while wearing protective equipment: lab gloves and coats.

Blood Conversion Kit Prototype

Safety is vital in the blood supply chain, especially as its products are ultimately transfused to patients. As such, in the design of our blood type conversion kit, we implemented a wide variety of safety measures. We devised a two-step blood type conversion kit to 1) mediate the enzymatic conversion process, and 2) purify converted RBCs in preparation for transfusion. We plan to pass washed RBCs through a series of airtight columns, using luer locks and biocompatible tubing, with bead-immobilized enzymes, where enzymatic conversion will take place. Our design was created with disposability in mind. Furthermore, we introduced post-conversion processing steps where we will isolate cleaved RBCs through the addition of antibodies and filtration with leukodepletion filters. We designed our kit after consultation with hematologists and feedback from blood banks, and visualized our hardware through Computer Aided Design. We verified the viability of various aspects of our kit design through a series of prototype experiments. In performing our proof of concept experiments, we elected to use porcine blood as an alternative to human blood, sourced from a farm in Hsinchu, a county in Taiwan, specializing in breeding pigs for scientific research purposes. With a reliable source and robust handling and screening procedures, we made sure our experiments were conducted in a safe manner.

References

US EPA. ATTACHMENT I--FINAL RISK ASSESSMENT OF ESCHERICHIA COLI K-12 DERIVATIVES, Feb. 1997.

Fred Hutchinson Cancer Research Center. “12.3 Corrosives.” Extranet, https://extranet.fredhutch.org/en/u/ehs/hamm/chap3/section12/subsection3.html