Difference between revisions of "Team:Freiburg/Project/System"

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The DiaCHIP is an innovative tool to screen for a broad range of antibodies present in blood serum. Antibodies can be an indicator for an immune response against an infection or a successful vaccination. They also play an important role in the diagnosis of autoimmune diseases. Especially the ability to differentiate between life threatening diseases and mild infections within a short time bears the potential to save lives.   
 
The DiaCHIP is an innovative tool to screen for a broad range of antibodies present in blood serum. Antibodies can be an indicator for an immune response against an infection or a successful vaccination. They also play an important role in the diagnosis of autoimmune diseases. Especially the ability to differentiate between life threatening diseases and mild infections within a short time bears the potential to save lives.   
 
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<b>Basic setup of the DiaCHIP</b>
  
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<b>Step 1: Basic setup of the DiaCHIP</b>
 
 
<div class="image_box left">
 
<div class="image_box left">
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<img align="left" src="https://static.igem.org/mediawiki/2015/5/55/Freiburg_generaloverview_RJ.jpeg" alt="Freiburg_generaloverview" width="400px">
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                                <div class="thumbinner">
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                                <a href="https://static.igem.org/mediawiki/2015/5/55/Freiburg_generaloverview_RJ.jpeg" class="lightbox_trigger">
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                                  <img src="https://static.igem.org/mediawiki/2015/5/55/Freiburg_generaloverview_RJ.jpeg" width="300px">
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                                                <div class="thumbcaption">
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                                                </a>
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                                                <p><B>Figure 1: DiaCHIP based on antigens derived from viruses and bacteria </B> weitere text.... </p>
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                                                </div>
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<div>
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The aim of our DiaCHIP is to screen simultaneously for hundreds of different infectious diseases. We based our system on antigens derived from viruses and bacteria (figure 1). If you get in contact with one of these diseases your immune system is producing antibodies. They are binding to the corresponding antigen. This binding event can be detected with our system. Our approach is based on two components. A silocone slide were the DNA coding for a distinct antigenic peptide is immobilized. The second component is a glas slide with a specific surface for the binding of the expressed antigens. Both are the size of a microscopy slide.
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see DNA Immobilization result LINK mit target blank
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<p>
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<b>Step 2: Cell-free</b>
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The DiaCHIP facilitates this process by a copying mechanism that converts a DNA template into a protein microarray by cell-free protein expression. This expression system based on a bacterial lysate makes the need for genetically engineered organisms to produce each single antigen redundant.
  
 
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<div class="image_box left">
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<img align="left" src="https://static.igem.org/mediawiki/2015/9/93/Freiburg_changeperspective.jpeg" alt="Freiburg_changeperspective" width="400px">
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                                  <img src="https://static.igem.org/mediawiki/2015/e/ee/Freiburg_overviewcellfree_RJ.jpg" width="300px">
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                                                </a>
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                                                <p><B>Figure 2: Cell-free.</B> </p>
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                                                </div>
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<div class="todo_box">
 
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Explain glass slide / PDMS sandwich
 
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Explain that antigens are immobilized on the glass slide and that they are detected with antibodies
 
 
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<b>Step 2: </b>
 
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<div class="todo_box">
 
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Explain that we have a label-free detection system
 
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<b>Step 3: </b>
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<p>
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<div class="todo_box">
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<b>Step 3: Glas surface</b>
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Explain that we fix DNA of the antigen epitopes on the PDMS slide
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</div>
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<img align="left" src="https://static.igem.org/mediawiki/2015/d/d5/Freiburg_systemoverview.png" alt="Freiburg_systemoverview" width="420px">
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                                <div class="thumbinner">
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                                <a href="https://static.igem.org/mediawiki/2015/7/79/Freiburg_specific_surface_RJ.jpg" class="lightbox_trigger">
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                                  <img src="https://static.igem.org/mediawiki/2015/7/79/Freiburg_specific_surface_RJ.jpg" width="300px">
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                                                <div class="thumbcaption">
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                                                </a>
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                                                <p><B>Figure 3: Surface.</B> </p>
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                                                </div>
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<b>Step 4: Preparing the DiaCHIP by Protein Synthesis</b>
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<b>Step 4: Measuring Serum Samples by iRIf</b>
 
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<div class="image_box left">
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Prior to screening for antibody-antigen interactions, the antigens have to be synthesized and immobilized in a microarray set-up. The DiaCHIP facilitates this process by a copying mechanism that converts a DNA template into a protein microarray by cell-free protein expression. This expression system based on a bacterial lysate makes the need for genetically engineered organisms to produce each single antigen redundant.
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</br>
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In order to collect the DNA templates, the respective sequences containing transcriptional and translational initiation sites, the antigen coding sequence and terminating regions have to be constructed and labeled with an amino group. An activated silicone slide provides the basis for immobilization of the DNA by covalent binding of the amino group. Spotting the antigen coding sequences in a distinct pattern enables to retrace a detected binding event to a certain disease.
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The template slide is placed in close proximity to the future protein array enabling the expressed proteins to reach the other surface by diffusion. We established a surface for specific immobilization of the target proteins. Thus, components of the expression mix can be washed away and do not hinder the analysis of the actual sample.
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                                <div class="thumbinner">
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                                <a href="https://static.igem.org/mediawiki/2015/5/56/Freiburg_iRiF_overview_RJ.jpg" class="lightbox_trigger">
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                                  <img src="https://static.igem.org/mediawiki/2015/5/56/Freiburg_iRiF_overview_RJ.jpg" width="300px">
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                                                <div class="thumbcaption">
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                                                </a>
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                                                <p><B>Figure 4: iRIf.</B> </p>
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                                                </div>
 
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<b>Step 5: Measuring Serum Samples by iRIf</b>
 
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</br>
 
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<!--korrigiert von Philipp05/09/15-->
 
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<!--neu von Ramona08/09/15-->
 
 
After preparation of the DiaCHIP, a patient’s serum sample can be flushed over the protein array. The binding of antibodies to the protein surface causes a minimal change in the thickness of the layer on the slide right at the corresponding antigen spot. This change can be measured without the need for a further label with an emerging method called iRIf (imaging Reflectometric Interference). Based on the interference of light beams reflected on different medium borders, binding events can be recorded in real-time.  
 
After preparation of the DiaCHIP, a patient’s serum sample can be flushed over the protein array. The binding of antibodies to the protein surface causes a minimal change in the thickness of the layer on the slide right at the corresponding antigen spot. This change can be measured without the need for a further label with an emerging method called iRIf (imaging Reflectometric Interference). Based on the interference of light beams reflected on different medium borders, binding events can be recorded in real-time.  
  
 
After weeks of optimizing the different components of the DiaCHIP, we are proud to present our results. We reached the highlight of our project with the successful <a href="https://2015.igem.org/Team:Freiburg/Results">detection of antibodies in our own blood!</a>
 
After weeks of optimizing the different components of the DiaCHIP, we are proud to present our results. We reached the highlight of our project with the successful <a href="https://2015.igem.org/Team:Freiburg/Results">detection of antibodies in our own blood!</a>
 
</p>
 
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</div>
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<p>
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<b>Step 5: Change perspective </b>
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<div class="image_box left">
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<div class="thumb2 trien" style="width:310px">
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                                <div class="thumbinner">
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                                <a href="https://static.igem.org/mediawiki/2015/9/93/Freiburg_changeperspective.jpeg" class="lightbox_trigger">
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                                  <img src="https://static.igem.org/mediawiki/2015/9/93/Freiburg_changeperspective.jpeg" width="300px">
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                                                <div class="thumbcaption">
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                                                </a>
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                                                <p><B>Figure 5: Change perspective.</B> </p>
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                                                </div>
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Revision as of 12:03, 16 September 2015

""

The DiaCHIP: Overview

When people click here, they know we are working on disease detection and antibodies, nothing specific!
On this page they need to learn about our DiaCHIP, at least enough to understand our results and be impressed. They need to be informed about:
- the glass slide / silicone sandwich (image + text)
- general workflow of the system
-> that we're working with epitopes of viruses/bacteria
- the basics of iRIf
- the "lets switch perspective" part of the presentation / the basics to understand the circles of the slide images used in the results section
try to think of how we explain the DiaCHIP in our presentation

RJ und JD kümmern sich drum

The DiaCHIP is an innovative tool to screen for a broad range of antibodies present in blood serum. Antibodies can be an indicator for an immune response against an infection or a successful vaccination. They also play an important role in the diagnosis of autoimmune diseases. Especially the ability to differentiate between life threatening diseases and mild infections within a short time bears the potential to save lives.
The DiaCHIP makes it possible to screen for multiple specific antibodies simply using a drop of blood.

Basic setup of the DiaCHIP

Figure 1: DiaCHIP based on antigens derived from viruses and bacteria weitere text....

The aim of our DiaCHIP is to screen simultaneously for hundreds of different infectious diseases. We based our system on antigens derived from viruses and bacteria (figure 1). If you get in contact with one of these diseases your immune system is producing antibodies. They are binding to the corresponding antigen. This binding event can be detected with our system. Our approach is based on two components. A silocone slide were the DNA coding for a distinct antigenic peptide is immobilized. The second component is a glas slide with a specific surface for the binding of the expressed antigens. Both are the size of a microscopy slide. see DNA Immobilization result LINK mit target blank

Step 2: Cell-free The DiaCHIP facilitates this process by a copying mechanism that converts a DNA template into a protein microarray by cell-free protein expression. This expression system based on a bacterial lysate makes the need for genetically engineered organisms to produce each single antigen redundant.

Figure 2: Cell-free.

Step 3: Glas surface

Figure 3: Surface.

Step 4: Measuring Serum Samples by iRIf

Figure 4: iRIf.

After preparation of the DiaCHIP, a patient’s serum sample can be flushed over the protein array. The binding of antibodies to the protein surface causes a minimal change in the thickness of the layer on the slide right at the corresponding antigen spot. This change can be measured without the need for a further label with an emerging method called iRIf (imaging Reflectometric Interference). Based on the interference of light beams reflected on different medium borders, binding events can be recorded in real-time. After weeks of optimizing the different components of the DiaCHIP, we are proud to present our results. We reached the highlight of our project with the successful detection of antibodies in our own blood!

Step 5: Change perspective

Figure 5: Change perspective.