Team:Bio Without Borders/Description

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Project motivation

We are from New York City and the Altantic Ocean is essentially next door. Horseshoe crabs are seen a lot throughtout the shores especially in Jamaica Bay. Unfortunately throughout the years the horseshoe crab population has been decreasing. We wondered why this was happening and after realizing what the pharmaceutical companies were doing with horseshoe crab blood. We wanted to find a different way in which we can optain the same protein that detects endotoxins without having to harvest bloood from horseshoe crabs.

Defining the project challenges

Even though the patent on Factor C assay has expired and new recombinant methods have been created — for example, Lonza bioscience has created an rFC assay — the assay still remains monopolized and inaccessible (in order to be able to view the Lonza protocol, you have to request permission from the company). It is also inaccessible in the sense that the protocol is very demanding in terms of the dollar amount, and requires expensive equipment and reagents. Therefore, the challenge of our project was to create a more accessible and easy-to-use diagnostic tool with the rFC in a way where we could visualize the self-cleavage of the protein.

How did we combat the challenges

We decided that our best course of action would be to create a second reporter protein, in our case a fusion protein between green fluorescent protein (GFP) and a cellulose binding domain (CBD), that included the cleavage site double Ile-Ile bond on the actual Factor C ,so that when the protein performed autocatalysis it would also recognize the site on the reporter and cleave that as well, causing a visual readout.