Prospects
How would we make Bacto-Aid?
Even though we spent hundreds of hours in the lab, we did not end up standing with the Bacto-Aid in our hands in Boston. But do not worry - we got a plan for how you can end up with our bandage. On this page we will enlighten you about the next steps in our project, which we would have followed if the days contained more than 24 hours or if we had a bunch of effective minions available. These steps have to be fulfilled before we can create a full functional patch that can be used in hospitals and in the everyday life.
Making a 4 monomer silk construct
There were some difficulties with producing a longer silk construct and whether it was due to the reproducibility of UCLA’s method or the too old streptavidin beads, we did not have time to conclude on. However, the first step would then of course be to try the ICA method with new streptavidin beads. When we have produced a 4 monomer silk construct, we would be inserting it into E. coli. and purify our silk from the bacteria. Here we would have liked to try a purification method without the use of His-tag, because earlier literature have demonstrated that recombinant silk produced without a His-tag have better mechanical properties compared to the fibres made from silk proteins with a His-tag Tokareva, O., et al. (2013). "Recombinant DNA production of spider silk proteins." Microbial biotechnology 6(6): 651-63.. For the spinning of silk we would first have tried to spin our silk by wet spinning, which is a very simple and inexpensive method Teulé, F., et al. (2013). "A protocol for the production of recombinant spider silk-like proteins for artificial fiber spinning." Nature protocols 4(3): 341-55.. In relation to using the recombinant silk for wound healing, it could also be interesting to prepare a three dimensional scaffold of the silk. Here a number of methods, such as salt leaching, gas forming or freeze-drying, have been reported to generate porous three-dimensional matrices Nazarov, R.., et al. (2004). "Porous 3-D Scaffolds from Regenerated Silk Fibroin" Biomacromolecules 5: 718-727.. The detailed description of how we want to proceed with a 4 monomer silk construct can be reviewed here.