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                 <h1>(2)Communicating science</h1>
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                 <h1>Outlook</h1>
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                 <p>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. </p>
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                 <h1>(3)Modeling cell-free expression</h1>
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                 <h1>Optical Detection: iRIf</h1>
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                 <p>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. </p>
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                 <p>One disadvantage of currently available serological tests is the need for secondary labels that allow the detection of disease markers. Making use of an optical method based on the interference of light, the DiaCHIP can detect specific binding events on a protein microarray without further labeling. Read more about this innovative tool and the physics behind it.  
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                <a href="https://2015.igem.org/Team:Freiburg/Project/iRIf" title="Optical Detection">iRIf Principle and Physics</a>
 
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                 <p> Want to read <a href="https://2015.igem.org/Team:Freiburg/Modeling">more</a>? </p>
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                 <h1>(4)Measuring our blood</h1>
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                 <h1>Surface Chemistry</h1>
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                 <p>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.</p>
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                 <p>The production of a customized protein microarray in the DiaCHIP is based on selective immobilization of antigens on a glass slide. Therefore, a specific surface chemistry needs to be established in order to reduce the proportion of unspecific binding of non-target proteins to a minimum.
 +
                  Read more about the different layer compositions we tested on our way to high specificity.
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                <a href="https://2015.igem.org/Team:Freiburg/Project/Surface_Chemistry" title="Surface Chemistry">Establishing a Specific Surface</a>
 
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                 <p> Want to read <a href="https://2015.igem.org/Team:Freiburg/Results">more</a>?</p>
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                 <h1>(5)DNA Engineering</h1>
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                 <h1>Protein Purification</h1>
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                 <p>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.
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                 <p>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 <i>E. coli</i> for being able to compare the results of both techniques. <br>
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                                <br>
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Read more about the overexpression and purification of several antigenic peptides. 
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Read more about how we combined different cloning methods to reduce our efforts in <a href="https://2015.igem.org/Team:Freiburg/Methods/Cloning" class="wikilink1">DNA Engineering.</a>
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                <a href="https://2015.igem.org/Team:Freiburg/Project/Protein_Purification" title="Protein Purification">Purification of Antigens</a>
 
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                 <h1>(6)Protein purification</h1>
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                 <p>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 <i>E. coli</i> for being able to compare the results of both techniques. <br>
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                 <p> A key feature of the DiaCHIP is the capability to produce protein arrays on demand via cell-free expression.  
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<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Project/Protein_Purification">Read more</a> about the overexpression and purification of several antigenic peptides. 
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                 <h1>(7)Surface chemistry</h1>
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                 <h1>DNA Engineering</h1>
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                 <p>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.<br>
+
                 <p>Genetic fusion of different antigens and tags is a basic requirement of our project. In order to enable several people to work in parallel we designed a cloning strategy easy to follow and additionally easy to expand for further needs.
−
<a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Project/Surface_Chemistry">Read more</a> about the different systems we tested on our way to high specificity.
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Read more about combining different cloning methods to reduce efforts in <a href="https://2015.igem.org/Team:Freiburg/Methods/Cloning" class="wikilink1">DNA Engineering.</a>
 
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                <a href="https://2015.igem.org/Team:Freiburg/Methods/Cloning" title="DNA Engineering">DNA Engineering</a>
 
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                 <h1>Diagnostics Today</h1>
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                 <p>One disadvantage of currently available diagnostic test is the need for secondary labels that allow the detection of disease markers. Making use of an optical method based on the interference of light, the DiaCHIP can detect specific binding events on a protein microarray without further labeling. <br>
+
                 <p>Currently used serological tests are available for a broad range of infectious diseases. However, they meet limitations that can be life-saving for some patient groups. The necessity of performing several tests in order to check for more than one disease at once is not only time-consuming but also costly. The DiaCHIP tackles these issues by providing a fast and affordable method for simultaneous testing.
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<a class=”wikilink1” href=”https://2015.igem.org/Team:Freiburg/Project/iRIf>Read more</a> about this innovative tool and the physics behind it.
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                <a href="https://2015.igem.org/Team:Freiburg/Design" title="Diagnostics today">Limitations and Solutions</a>
 
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Revision as of 18:48, 17 September 2015

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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 chip 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 as indicator for an immune response towards an infectious diseases or a successful vaccination. They also play an important role in the diagnosis of autoimmune diseases. Identifying diseases by detecting disease associated 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 of disease related antigens and a novel method for label-free detection - all this 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.

  • The DiaCHIP - System Overview

    The core of our new diagnostic device are two slides that form a microfluidic chamber. Therein an antigen array can be generated on demand on the bottom slide. By flushing the chamber with a blood sample, binding of antibodies present in the sample is detected with the optical detection method iRIf in real-time.

    Step by Step Overview

  • Outlook

    Want to read more?

  • Optical Detection: iRIf

    One disadvantage of currently available serological tests is the need for secondary labels that allow the detection of disease markers. Making use of an optical method based on the interference of light, the DiaCHIP can detect specific binding events on a protein microarray without further labeling. Read more about this innovative tool and the physics behind it.

    iRIf Principle and Physics

  • Surface Chemistry

    The production of a customized protein microarray in the DiaCHIP is based on selective immobilization of antigens on a glass slide. Therefore, a specific surface chemistry needs to be established in order to reduce the proportion of unspecific binding of non-target proteins to a minimum. Read more about the different layer compositions we tested on our way to high specificity.

    Establishing a Specific Surface

  • 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 several antigenic peptides.

    Purification of Antigens

  • Cell-Free Expression

    A key feature of the DiaCHIP is the capability to produce protein arrays on demand via cell-free expression.

    Want to read more?

  • DNA Engineering

    Genetic fusion of different antigens and tags is a basic requirement of our project. In order to enable several people to work in parallel we designed a cloning strategy easy to follow and additionally easy to expand for further needs. Read more about combining different cloning methods to reduce efforts in DNA Engineering.

    DNA Engineering

  • Diagnostics Today

    Currently used serological tests are available for a broad range of infectious diseases. However, they meet limitations that can be life-saving for some patient groups. The necessity of performing several tests in order to check for more than one disease at once is not only time-consuming but also costly. The DiaCHIP tackles these issues by providing a fast and affordable method for simultaneous testing.

    Limitations and Solutions