Safety

Overview

Throughout our project, we obeyed the iGEM safety rules. When conducting our experiments, all of the organisms used for developing our product stayed in our lab. To ensure safety during future use of our device outside the lab, we made sure that no biomaterials were spread to the environment.

Laboratory Safety

Safety Training inside the Laboratory

Before we entered the lab, all of our team members had taken the "Laboratory Biosafety Training" course, "Laboratory Safety & Hygiene Training" course and “Hazard Communication Training” course and received respective certificates from NYCU. Furthermore, all experiments were strictly examined by the NYCU laboratory management system to ensure all procedures were safe.

Level and Rule of the laboratory

Our experiments were done in a BSL1 laboratory. Also, when we performed experiments in the laboratory, our advisers provided training regarding the correct operation of instruments, methods preventing containment, and ways to deal with laboratory waste. What's more, they supervised us during our experiments to ensure safety. All equipment used to experiment with and contain Escherichia coli (E. coli) were sterilized with alcohol or bleach properly, and experiments were operated in Laminar flow hood to prevent any leakage or pollution of bacteria. We familiarize ourselves with and follow Taiwan biosafety laws.

Safety of UBX Product

Processing Procedure in the Lab

In the future, we plan to utilize our product in medical, agricultural, industrial fields, etc. Therefore, we should guarantee that Ubx is safe to humans and harmless to the environment. As a result, we design a processing standard in the laboratory.

1. Free of E. coli

The fiber should not contain any genetically-modified organisms. Therefore, after we expressed Y167 mRFP Y240 with E. coli, which was sonicated by the sonicator to let protein be soluble in the Tris Base solution. In the crosslinking reaction step, we mixed the sonicated Tris lysate with EDTA-Fe solution, and then added H₂O₂ into the protein mixture to initiate the reaction. In order to verify the lysate is free of E. coli, we spread it on the LB agar plate, and incubated at 37°C for 16 hours. The results show no living E. coli on the plate.

2. Harmless to human body

Since the Y167 Y240 regions are separated from Ubx protein, it’s essential to show that Ubx protein is harmless to the human body.

Ubx materials, unlike elastin and collagen, are not naturally found in vivo. It is particularly important to determine whether Ubx materials are biocompatible to evaluate their potential for in vivo applications.

According to the paper, Ubx fibers were cultured with three primary human cell lines derived from vasculature: umbilical vein endothelial cells, brain vascular pericytes, or aortic smooth muscle cells. Cells readily adhered to Ubx fibers, and could be transferred between tissue cultures without loss of viability for at least 96 hours.1

Furthermore, Ubx fibers cultured with macrophages in vitro did not lyse or activate the macrophages, and did not cause hemolysis when incubated with human red blood cells.2

Since Ubx materials are cytocompatible, nonimmunogenic, noninflammatory, and nonhemolytic, we consider that the materials of Y167 Y240 regions from Ubx used for medical applications must demonstrate excellent biocompatibility and no need to worry about whether it will harm the human body.