Solution: Assays using human tissue culture / ELISA assays

In testing whether our insulin produced was the correct structure and functioned in the correct way, we decided to avoid performing a mouse convulsion test which is the standard method used. In order to avoid the ethical issues surrounding induced murine fatality, we decided to test our insulin using an ELISA assay with an antibody specific to human insulin. This would determine whether our proinsulin had the correct three-dimensional structure.

As our single chain insulin will have a different structure, we do not expect it to interact with an insulin specific antibody. We will further test our insulin performing assays on human tissue culture – adipocyte tissue and/or pancreatic tissue. These assays were performed alongside insulin manufactured from different pharmaceutical companies to compare the response. These two assays will prevent the need to use live biological organisms. Importantly, using human cell lines will allow us to determine directly whether our single-chain insulin and proinsulin interacts with the correct insulin receptor in stimulating the correct response. An adipocyte assay will thus provide us with the same results as a mouse convulsion test without the same ethical issues.

Issue: Insulin can be fatal when injected

Solution: prevent handling of any needles and handling of small amounts

As our biological parts that are constructed involves plasmids without any potential infectious particles eg. viral particles, we expect shipping to be very safe. Our constructs itself are already placed in the pSB1C3 vector that is standard in the iGEM registry, without a replicative origin. In order to prevent any additional issues, we also ensured all biological materials had double containment – containment in eppendorf tubes, additionally sealed using parafilm and/or clingwrap.

Specific to the experience of our laboratory, we found that in Australia our strict border control provided us with difficulties in obtaining our parts in the mail. With a bit of foresight and open communication, we could potentially mitigate this issue for the iGEM HQ.

Before starting any experimental work, the entire USYD iGEM 2017 team performed extensive safety research and inductions which included reading and signing off on relevant safety forms. As the laboratory space that we used a Physical containment Level 2 (PC2), safety procedures were very strict and performed by our supervisor Nick Coleman. The induction involved direction to all the relevant points of emergency eg. the fire blanket, fire escape, shower, ’eye wash basin lol’, location of all flammable goods, hazardous waste disposal and also designated areas for handling specific chemicals like acrylamide and bromothymol blue.

In addition to always wearing laboratory coats and latex gloves at all times, our laboratory also introduced compulsory wearing of safety glasses.


Our safety module involved an additional online component

addressing occupational health and safety hazards. This came standard with the University of Sydney requirements and involved presenting the participant with hypothetical situations with associated risks. We then had to identify and rank the dangers that arose in the situations. This provided all the laboratory workers with adequate information and identified potential risks before they happened. This procedure is standard in USYD.

An addition to laboratory safety, our team was also well versed in general laboratory etiquette including cleaning of used equipment to prevent any cross contamination and maintaining the areas we worked in (particularly with any staining and any handling of bacterial cultures). This was extremely important since the laboratory we worked in included other microorganisms and viral particles and containment was extremely important. Any issues experienced were raised, discussed and solved in weekly meetings in conjunction with other honours and PhD students from the laboratory.