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Results

Results

Here you can find all the results of our experiments, the conclusions that can be drawn from them, and the outlook for future research

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In an attempt to impact the health sector and improve the quality of life of patients suffering from inflammatory bowel disease (IBD), cELPro was designed. As is described in more detail in the Project Description, the purpose of this therapy is to make treatment of IBD more affordable, more specific, and less invasive. To this end, we created genetically engineered bacteria, which were inhibited from dividing due to in situ hydrogelation from block Elastin-Like Polypeptides (ELPs). In addition, they were still able to produce a target protein such as a therapeutic protein, IL-10, for the treatment of IBD or mNeonGreen, a model protein. This platform could be less infectious while still retaining the ability to produce drugs for the treatment.

Achievements

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We successfully cloned the genetic sequences encoding the self-assembling ELPs and expressed them for the designated hydrogel at different weight-to-volume percentages.

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We managed to form this hydrogel inside the bacteria which made them more resistant to ampicillin and furthermore inhibited the division of these cells, making them more controllable.

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Finally, we were able to express an ELP-mNeonGreen fusion protein inside the bacteria after formation of the hydrogel, indicating that the bacteria are still functional after intracellular gelation.

Scroll down below to find a more elaborate reporting and discussion of the results from the different stages of the project.

Cloning and Expression of ELPs

To make our bacteria more controllable we wanted to prevent them from dividing. We designed a sequence of ELPs that are able to form a hydrogel within the cells, and potentially inhibit bacteria division. We cloned these sequences into the appropriate vectors, expressed the proteins and subsequently characterized them.

Hydrogel

In this section, we show that we were able to form hydrogels of our constructs. We then test the properties and utilities of these gels with a variety of experiments. These include testing the diffusion rates, viability tests of the gelated bacteria and microscopy.

Therapeutic

As a proof of concept, we decided to express a therapeutic for the treatment of IBD. After speaking to several experts and stakeholders, and doing our own literature research, we found the cytokine IL-10 to be the best candidate for this purpose. Apart from expressing the protein, we tried to perform an immunoassay on it in order to quantify the expression levels.

Outlook

Our project has known its successes and failures. Both mean that there is still enough work that could be done to improve the platform. Here, we provide an overview of what we think the right next steps are for the project. We think that, with the right optimization, there is a lot of potential for the platform.

If the boxes to the result pages do not appear, please click the links here:

Cloning

Hydrogel

Therapeutic

Outlook

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