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Difference between revisions of "Team:CAU China/Design"

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<h3>Reducing Toxin Levels</h3>
  
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          In an attempt to push our purity towards enabling intravenous administration, we decided to perform fractionation in a pressure-driven size-exclusion filter system. Suitability of the phage product for intravenous application could then be assessed with an additional endotoxin test.
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          After these steps, our phages would be ready for animal testing, which we did not plan to attempt in the context of the iGEM competition
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  <h2>Encapsulation</h2>
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        <The goal of this module is to optimize the packaging of assembled phages to prepare them for
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        application to patients.>
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          The transport within the body often poses problems for the activity and integrity of phages, which can be addressed by packaging of the phages in a protective layer<sup><a href="#phareferences">5</a></sup>. For oral application of our phage product, sufficient stability for gastric passage has to be guaranteed. The highly acidic environment, as well as the presence of proteases composed the major challenges. Our packaging method thus needed to provide resistance to low pH. At the same time, phages needed to be released from their protection upon reaching intestinal fluid. These characteristics are provided by calcium-alginate microspheres<sup><a href= https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5264180/>6</a></sup>. Lacking suitable encapsulation hardware, we decided to build our own nozzle to encapsulate the phages in monodisperse microcapsules.
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        </p>
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        <p>
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          The methods most suitable for quantification of size and assessment of monodispersity of the alginate capsules were brightfield and epifluorescence microscopy. To determine whether our encapsulation method fulfills our requirements of survival and release of phages in simulated gastric and intestinal fluid, respectively, we subsequently performed plaque assays.
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Revision as of 02:42, 19 June 2019










Phactory

Project Design

The Problem

Alice was beginning to get very tired of sitting by her sister on the bank, and of having nothing to do: once or twice she had peeped into the book her sister was reading, but it had no pictures or conversations in it, 'and what is the use of a book,' thought Alice 'without pictures or1There was nothing so VERY remarkable in that; nor did Alice think it so VERY much out of the way to hear the Rabbit say to itself, 'Oh dear! Oh dear! I shall be late!' (when she thought it over afterwards, it occurred to her that she ought 2 This was the overarching problem our project needed to tackle.

Either the well was very deep, or she fell very slowly, for she had plenty of time as she went down to look about her and to wonder what was going to happen next.

fore seen a rabbit with either a waistcoat-pocket, or a watch to take out of it, and burning with curiosity, she ran across the field after it, and fortunately was just in time to see it pop down a large rabbit-hole under the hedge.

Reducing Toxin Levels

In an attempt to push our purity towards enabling intravenous administration, we decided to perform fractionation in a pressure-driven size-exclusion filter system. Suitability of the phage product for intravenous application could then be assessed with an additional endotoxin test.

After these steps, our phages would be ready for animal testing, which we did not plan to attempt in the context of the iGEM competition

Encapsulation

The transport within the body often poses problems for the activity and integrity of phages, which can be addressed by packaging of the phages in a protective layer5. For oral application of our phage product, sufficient stability for gastric passage has to be guaranteed. The highly acidic environment, as well as the presence of proteases composed the major challenges. Our packaging method thus needed to provide resistance to low pH. At the same time, phages needed to be released from their protection upon reaching intestinal fluid. These characteristics are provided by calcium-alginate microspheres6. Lacking suitable encapsulation hardware, we decided to build our own nozzle to encapsulate the phages in monodisperse microcapsules.

The methods most suitable for quantification of size and assessment of monodispersity of the alginate capsules were brightfield and epifluorescence microscopy. To determine whether our encapsulation method fulfills our requirements of survival and release of phages in simulated gastric and intestinal fluid, respectively, we subsequently performed plaque assays.


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