Safety
Overview

Our PolYneer project aims to create a novel bispecific antibody by using the synthetic biology method, that targets the c-MET and EGFR mutation simultaneously in the resistance situation that was created after treating Osimertinib for NSCLC patients.

To ensure a safe and secure working environment, we are committed to identifying and managing potential risks continuously. Our goal is to minimize hazards to our iGEM team, university colleagues, society, and the environment. Our safety approach takes into consideration various factors, including project design safety and safety protocols in the laboratory. In the following sections, we will outline the safety measures we have implemented throughout the design and experimental phases.

General Safety
Figure 1 Laboratory environment
Figure 2 Laboratory environment
Safety in Project Design

Cloning

During our project, different genes were assembled for cloning into various E. coli bacterial strains, including TOP10 and BL21(DE3). These strains are non-pathogenic to humans and pose no hazards to the environment. They also fall within the White List.

Genes

We have designed and expressed six composite parts through the bacterial system to produce a component of our bispecific antibody. None of these parts are associated with the production of toxic substances.

Used genes and their risk associated.
Part's nameassociated
eGFP-Ssp-Mxe inteinNo proved hazardous effect
Peptide-Ssp-Mxe inteinNo proved hazardous effect
Scrb peptide-Ssp-Mxe inteinNo proved hazardous effect
eGFP-Npu-SsrA inteinNo proved hazardous effect
Peptide-Npu-SsrA inteinNo proved hazardous effect
Scrb peptide-Npu-SsrA inteinNo proved hazardous effect
Table 1 Used genes and their risk associated.

Mammalian Cell Lines

In all of our mammalian cell work, we utilized HCC827 and NCL-H1975 cell lines. These cell lines are classified as BSL-1 materials and have undergone testing and certification to ensure they are free of known pathogens. Hence, both cell lines are included in the White List.

Safety Laboratory Work

Prior to commencing any experiments, the team underwent thorough safety training to ensure a secure working environment. Throughout our time in the lab, we had the invaluable support of a dedicated safety officer who was readily available to address any inquiries or concerns we might have had. To guarantee the utmost precaution, all our protocols underwent rigorous scrutiny by experienced professors and advisors before implementation. Continuous supervision by these knowledgeable individuals, as well as other researchers in the lab, was diligently maintained. This collective commitment to safety ensured that we operated within the highest standards throughout our experimental endeavors.

Our Safety Practices

Appropriate and required protection was used during all steps of wet lab manipulations.

ExperimentSafety precautions
Handling of Gelred for electrophoresis analysisEnsuring the glove is intact and equipped to prevent direct contact with skin
Observing the gel band under UV boxWearing UV protective goggles or an UV face shield to prevent prolonged exposure to UV radiation
Sterile experiment inteinRemoving all flammable materials approaching the Bunsen Burner
Handling acids and corrosive chemicalsAllocating them in fumehood delicately
Table 2Among protocols that required a special safety consideration

Laboratory Security

The security rules, and good practices on lab and security materials are listed below:

General Laboratory Safe Practices

Assess risks from your laboratory activities and implement proper safety measures

Establish Safe Operating Procedure (SOP)

Registered and passed safety training from Health and Safety Office (HSO)

Conduct regular self-inspection

Conduct regular housekeeping check

With the help of our lab security manager, we have listed chemical products which need special attention during manipulation:

Hydrochloric acid (HCl)Corrosive
GelredHarmful, irritant
PolyacrylamideCarcinogenic and mutagenic
SDSFlammable
EthanolFlammable
KanamycinToxic