Team:TU Eindhoven/Project

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        <span>Introduction</span>   
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      <p class="para">This year’s iGEM team of the Eindhoven University of Technology focuses on a fundamental problem in the application of genetically modified bacteria inside the human body – the immune system. Locally bacterial produced drugs are a promising future in the field of medical treatments. However, this local drug release and production requires bacterial life forms inside the human body and these bacteria can cause immune responses. This natural immune system can be evaded with the use of two methods: suppression of the entire immune system or modifying the used bacteria in order to minimise the immune systems’ response. Due to the devastating impact of the first possibility, iGEM 2014 team Eindhoven decided to proceed with the second option.  </p>
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      <p class="para">This year’s iGEM team of the Eindhoven University of Technology focuses on a fundamental problem in the application of genetically modified bacteria: bacteria are not suited for many environments.
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For example, local bacterial production of medicine has a promising future as a medical treatment, but the human immune system is still a big limiting factor for this new technology. Another field that would greatly benefit from more resilient bacteria is the bioreactor industry. Bacteria that can survive in high pressure and temperature environments can help increasing efficiency of reactors.
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To create such resilient bacteria the 2014 team has designed a ‘plug and play’ system using copper free click chemistry to attach different chemical groups to create bio-layers on E. Coli cell membranes. Circularly permuted OmpX (CPX), an outer membrane protein, was mutated to contain an azido-functionalized unnatural amino acid. CPX functions as an anchor for any DBCO functionalized molecule to click onto. The polymers used in this project were designed to form hydrogels, which enables the bacteria to have antifouling properties.
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  <span>Current solution</span>   
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  <span>Application</span>   
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      <p class="para">Bacteria can be made undetectable to the immune system with the use of encapsulation. Hydrogels are suitable for this purpose. Hydrogels are water-absorbing (synthetic) polymers, and are therefore able to form a layer around the bacteria. Due the low reactivity of this hydrogel capsule towards the immune system and permeability to small essential molecules (for example nutrients and wastes). With the use of microfluidic techniques, the amount of bacterial cells per liquid droplet is relatively easy to control. In the produced microfluidic droplet, hydrogel formation can be induced. The result is a hydrogelation from the outside towards the core of the droplet, surrounding the entire group of bacteria. The problems with these current techniques are: inhomogeneous hydrogelation (due to gelation form the outside to the core), uncontrollable cell growth inside the encapsulation, thus incontrollable drug release, and non-degradable encapsulations. The conventional encapsulation technique is visualised in Figure 1. </p>
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      <p class="para">With our team’s biomedical background in mind, an anti-fouling chemical layer for use in the human body was chosen to test the ‘plug and play’ system. An anti-fouling hydrogel has to have little to no interaction with the human immune system, thus preventing immune responses caused by the presence of bacteria. Dibenzocyclooctyne Polyethylene glycol 10kDa (DBCO-PEG 10kDa) was chosen as the molecule to click onto OmpX to form the hydrogel because it has good anti-fouling properties and the modular length allows for easy testing on a smaller scale.</p>
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  <span>Microfluidics</span>   
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      <p class="para">In order to precisely control the hydrogel formation, microfluidic devices are used in which the conditions are optimal to form individually encapsulated cells. This way clustering of cells is prevented and the end product will be usable beads instead of large aggregated blobs.  </p>
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Latest revision as of 13:22, 12 September 2014


Project Description

Project Description

Introduction

This year’s iGEM team of the Eindhoven University of Technology focuses on a fundamental problem in the application of genetically modified bacteria: bacteria are not suited for many environments. For example, local bacterial production of medicine has a promising future as a medical treatment, but the human immune system is still a big limiting factor for this new technology. Another field that would greatly benefit from more resilient bacteria is the bioreactor industry. Bacteria that can survive in high pressure and temperature environments can help increasing efficiency of reactors.

To create such resilient bacteria the 2014 team has designed a ‘plug and play’ system using copper free click chemistry to attach different chemical groups to create bio-layers on E. Coli cell membranes. Circularly permuted OmpX (CPX), an outer membrane protein, was mutated to contain an azido-functionalized unnatural amino acid. CPX functions as an anchor for any DBCO functionalized molecule to click onto. The polymers used in this project were designed to form hydrogels, which enables the bacteria to have antifouling properties.



Application

With our team’s biomedical background in mind, an anti-fouling chemical layer for use in the human body was chosen to test the ‘plug and play’ system. An anti-fouling hydrogel has to have little to no interaction with the human immune system, thus preventing immune responses caused by the presence of bacteria. Dibenzocyclooctyne Polyethylene glycol 10kDa (DBCO-PEG 10kDa) was chosen as the molecule to click onto OmpX to form the hydrogel because it has good anti-fouling properties and the modular length allows for easy testing on a smaller scale.



Microfluidics

In order to precisely control the hydrogel formation, microfluidic devices are used in which the conditions are optimal to form individually encapsulated cells. This way clustering of cells is prevented and the end product will be usable beads instead of large aggregated blobs.

Created by iGEM Team TU Eindhoven 2014

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