Team:Freiburg/Project/Overview

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The DiaCHIP - A Versatile Detection System

DiaCHIP_Sabi

Project Motivation

Serological tests are a key element in modern medicine. Especially for detection and identification of infectious diseases, performing several blood tests is inevitable. Testing for more than one disease at a time or diagnosing a patient with diffuse symptoms usually requires more than one serological test. Every single test that is performed increases the time of waiting for a result as well as the bill. In case of dangerous infectious diseases every minute until the onset of the appropriate treatment is important for life. What if there was a possibility to combine all this testing in one single device which is affordable for everyone?

Detecting Antigen-Antibody Interactions

The DiaCHIP is an innovative tool to screen for a broad range of antibodies present in serum. Antibodies serve indicator for an immune response towards an infection or a successful vaccination. They also play an important role in the diagnosis of autoimmune diseases. Spotting diseases by detecting correspondent antibodies in a patient's serum is an established method in modern diagnostics.
Based on the very same principle, the DiaCHIP enables to simultaneously screen for multiple diseases at time, thereby reducing time and cost of a diagnosis. Especially the ability to differentiate between life threatening diseases and mild infections within a short time bears the potential to save lives.

The Concept

The key feature of the DiaCHIP concept is the combination of on-demand protein synthesis and a novel method for label-free detection packed into one device. The idea is to overcome challenges commonly found in protein array production and preservation. In addition, results can be obtained in a time- and cost-efficient manner; with a device simple enough to be rebuilt by future iGEM Teams.

  • Building our own device

    The device originally used, in collaboration with AG Roth, is an expensive machine based on rather simple components. Therefore, we decided to build our apparatus in a cost-efficient manner. We were able to produce reliable results with it and provide a construction plan. This plan will make it possible for future iGEM generations to build and use their own label-free protein array analysis tool.

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  • Communicating science

    Diagnosing diseases fast and reliable is not only an issue among medical staff, it is also subject to public interest. This has lead us to ask for people's opinions regarding the DiaCHIP. Although the method is based on synthetic biology, which is a problematic term for the broad public according to a survey initiated by the Leopoldina (National academy of science), we received a lot of positive feedback.

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  • Modeling cell-free expression

    In order to optimize the DiaCHIP for future applications, we optimized the process of cell-free expression and diffusion over time. Making use of xxx parameters and xxx ordinary differential equations, we computed the size of the resulting antigen spots on the protein array and identified the factors limiting cell-free expression in the DiaCHIP.

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  • Measuring our blood

    One of the most promising results was obtained from the detection of anti-tetanus antibodies in human blood serum. The DiaCHIP analysis made it possible for us to distinguish serum samples from a team member before and after vaccination. Samples taken two weeks after vaccination produced higher signals, compared to those prior to antigen exposure.

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  • DNA Engineering

    Genetic fusion of different antigens and tags is a really basic requirement of our project. To enable many people to work on our DNA constructs in parallel we designed a cloning strategy easy to follow and additionally easy to expand for further needs.
    Read more about how we combined different cloning methods to reduce our efforts in DNA Engineering.

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  • Protein purification

    Protein expression in the DiaCHIP is mediated by cell-free expression. As this is an advanced method dependent on the optimization of many parameters, we got back to conventional protein purification in E. coli for being able to compare the results of both techniques.
    Read more about the overexpression and purification of many antigenic peptides.

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  • Surface chemistry

    The production of a protein microarray in the DiaCHIP is based on specific immobilization of antigenic peptides on a glass slide. Therefore, we optimized a protocol for a specific surface chemistry until the proportion of unspecific binding of non-target proteins was reduced to a minimum.
    Read more about the different systems we tested on our way to high specificity.

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  • Measuring our blood

    One of the most promising results was obtained from the detection of anti-tetanus antibodies in human blood serum. The DiaCHIP analysis made it possible for us to distinguish serum samples from a team member before and after vaccination. Samples taken two weeks after vaccination produced higher signals, compared to those prior to antigen exposure.

    Want to read more?