Difference between revisions of "Team:Freiburg/testpage"

Line 1: Line 1:
−
{{Freiburg/CSS}}
+
{{Freiburg/test}}
 
{{Freiburg/MenubarTest}}
 
{{Freiburg/MenubarTest}}
−
{{Freiburg/wiki_content_start}}
 
−
<!-- wiki content goes in here -->
 
  
 +
{{Freiburg/wiki_content_start}}
 
<html>
 
<html>
  
 
<style>
 
<style>
 
/*========= BEGIN: style for navigation bar ==========*/
 
/*========= BEGIN: style for navigation bar ==========*/
−
#results {
+
#project {
−
     background: url(https://static.igem.org/mediawiki/2015/2/2e/Freiburg_icon_results_active_yellow.png) no-repeat;
+
     background: url(https://static.igem.org/mediawiki/2015/d/da/Freiburg_icon_project_active_yellow.png) no-repeat;
 
}
 
}
  
−
#results a {
+
#project a {
 
     color: #ecdc18;
 
     color: #ecdc18;
 
}
 
}
  
 
#runningchip {
 
#runningchip {
−
     left: 52%;
+
     left: 18.6%;
 
}
 
}
 
/*========= END: style for navigation bar ==========*/
 
/*========= END: style for navigation bar ==========*/
  
 +
/* code adapted from http://www.smashingmagazine.com/2012/04/pure-css3-cycling-slideshow/*/
  
−
#tetanus_leftside, #agfp_leftside {
+
#slider {
−
padding-right:30px;
+
  background: #FCFCFC;
 +
  height: 500px;
 +
  width: 900px;
 +
  overflow: visible;
 +
  position: relative;
 
}
 
}
−
#tetanus_leftside p, #agfp_body p, #device_results_body p{
 
−
    color: #000;
 
−
font-size: 20px;
 
−
line-height: 30px;
 
−
word-spacing: 0px;
 
  
 +
/* hides everything outside the slider box */
 +
#mask {
 +
  overflow: hidden;
 +
  height: 500px;
 
}
 
}
−
#tetanus_results{
+
 
−
clear: both;
+
/* initialize position for positioning slides outside the slider */
 +
#slider ul {
 +
  margin: 0;
 +
  padding: 0;
 +
  position: relative;
 
}
 
}
−
#tetanus_rightside, #agfp_rightside{
+
 
−
float:right;
+
/* define image properties and position them outside the slider mask */
−
max-width: 450px;
+
#slider li {
−
color: #000;
+
  width: 900px; /* Width slide */
 +
  height: 500px; /* Height slide */
 +
  position: absolute;
 +
  right: 0; /* Original Position - Outside of the Slider */
 +
  list-style: none;
 
}
 
}
  
−
#tetanus_leftside, #agfp_leftside{
+
 
−
max-width: 380px;
+
#first,
 +
#second,
 +
#third,
 +
#fourth{
 +
  left: -1000px;
 +
  -ms-transition: tranform 1s ease;
 +
  -webkit-transition: transform 1s ease;
 +
  transition: transform 1s ease;
 +
  font-size: 100%;
 
}
 
}
  
−
#tetanus_rightside{
+
#first{left: 0;}
 +
#third{z-index : -100;}
 +
#fourth{z-index: -100; left: 1000px;}
  
 +
.container{
 +
    position: relative;
 +
    height: 500px;
 +
    margin: 3% 0;
 +
    width: 1000px;
 
}
 
}
  
−
h1, h2{
+
#buttonleft {
−
font: "Roboto";
+
  z-index: 200;
−
font-size: 30pt;
+
  position: absolute;
−
color: #000000;
+
  top: 190px;
−
font-weight: bold;
+
  left: 0;
−
line-height: 1.3;
+
 
}
 
}
−
#tetanus_result_image_leading{
+
 
−
padding-top: 20px;
+
#buttonright{
−
padding-bottom: 15px;
+
  z-index: 200;
−
color: 000;
+
  position: absolute;
−
font-weight: bold;
+
  right: 0;
−
font-size: 15pt;
+
  top: 190px;
−
line-height: 1.3;
+
 
}
 
}
−
#tetanus_result_image{
+
 
−
margin-left: auto;
+
#buttonleft img,
−
margin-right: auto;
+
#buttonright img{
−
width: 450px;
+
  width: 50px;
 
}
 
}
  
−
#tetanus_result_image img{
+
#content-slider{
−
width: 450px;
+
  position: absolute;
 +
  top:0;
 +
  left:50px
 
}
 
}
  
  
−
#agfp_results{
+
#buttonright:hover,
−
clear: both;
+
#buttonleft:hover{
−
padding-top:75px;
+
  cursor: pointer;
 +
  cursor: hand;
 
}
 
}
−
#agfp_leftside{
+
 
−
width:380;
+
.container{
−
max-width: 380px;
+
  position: relative;
−
margin-left:auto;
+
−
margin-right:auto;
+
 
}
 
}
−
#agfp_leftside img{
+
 
−
display:inline;
+
.artboard{
−
align: center;
+
    box-sizing: border-box;
−
width: 380px;
+
    height: 470px;
−
height: 510px;
+
    border-radius: 60px;
−
margin-left:auto;
+
    background-color: rgba(0, 81, 162, 0.3);
−
margin-right:auto;
+
}
−
}
+
 
−
#agfp_body{
+
.sliderimage{
−
padding-top: 25px;
+
  float: right;
 +
  margin: 3% 75px 3% 25px;
 +
  width: 350px
 
}
 
}
  
−
#device_results{
+
.slidertext{
−
clear: both;
+
  margin: 3% 25px 0 75px;
−
padding-top:75px;
+
  width: 350px;
 +
  text-align: justify;
 +
  line-height: 1;
 +
  float: left;
 
}
 
}
−
#device_results_headline{
+
 
−
text-align: center;
+
.slidertext h1{
−
}
+
  text-align: left;
−
#device_results_image{
+
−
padding-top: 30px;
+
−
margin: auto;
+
−
width:400px;
+
−
}
+
−
#device_results_image img{
+
−
+
−
width: 400px;
+
−
}
+
−
#device_results_body{
+
−
padding-top: 30px;
+
 
}
 
}
  
−
@media screen and (min-width: 963px){
+
.slidertext p{
−
    #agfp_leftside,  #tetanus_leftside{
+
margin-bottom: 0;
−
        float: left;
+
−
    }
+
−
#tetanus_rightside, agfp_rightside{
+
−
max-width: 450px;
+
−
}
+
−
#tetanus_leftside, agfp_leftside{
+
−
max-width: 380px;
+
−
}
+
−
+
−
    #agfp_rightside,  #tetanus_rightside{
+
−
        float: right;
+
−
    }
+
 
}
 
}
  
−
@media screen and (max-width: 962px){
+
.slidertext.indent{
−
    #agfp_leftside,  #tetanus_leftside, #agfp_rightside,  #tetanus_rightside{
+
  clear: both;
−
        float: none;
+
  margin-left: 15%;
−
        width: auto;
+
  margin-top: 0;
−
max-width:none;
+
  width: 290px;
−
min-width:none;
+
−
margin-left: auto;
+
−
margin-right: auto;
+
−
 
+
 
}
 
}
  
−
h2{
+
.dummy-image{
−
color: #EEE;
+
  background-color: black;
−
font-size: 200%;
+
  height: 400px;
 +
  width: 350px;
 
}
 
}
  
Line 152: Line 156:
  
  
 +
<html>
  
−
<div class="content_box">
+
<script>
 +
$(document).ready(function(){
  
−
<div id="anti_tetanus_results">
+
    var distance = 2;
−
<div class="floatbox right">
+
    // see: http://atomicrobotdesign.com/blog/web-development/how-to-disable-a-button-until-a-jquery-animation-has-finished/ //
−
<h2>
+
    // bind the button to the function, so it can be unbound in the code and the button isnt clickable any more //
−
We successfully diagnosed anti-Tetanus antibodies in human blood serum
+
    $("#buttonleft").bind("click", moveleft);
−
</h2>
+
function moveleft(e) {
−
<p>
+
// unbind the button //
−
Check out this video showing how the antibodies in the serum bind our tetanus antigens on the DiaChip
+
$("#buttonleft").unbind();
−
</p>
+
console.log("Click "+distance);
−
<div class="image_box right">
+
−
<img src="https://static.igem.org/mediawiki/2015/7/74/Freiburg_labjournal-roi_selection_20150817_429.png" width="100%"></img>
+
−
</div>
+
−
</div>
+
  
 +
if (distance == 1) {
 +
console.log("Distance: "+distance);
 +
$('#first').css({"transform": "translateX(0)", "-webkit-transform": "translateX(0)", "-ms-transform": "translateX(0)"});
 +
    $('#second').css({"z-index": "100"});
 +
    $('#third').css({"z-index": "-100", "transform": "translateX(0)", "-webkit-transform": "translateX(0)", "-ms-transform": "translateX(0)"});
 +
    $('#fourth').css({"transform": "translateX(0)", "-webkit-transform": "translateX(0)", "-ms-transform": "translateX(0)"});
 +
    distance = 2;
 +
}
  
−
<div class="floatbox left">
+
else if (distance ==2) {
−
<p>
+
console.log("Distance: "+distance);
−
We took a persons blood before and after a tetanus vaccination and screened for a tetanus infection.  
+
$('#first').css({"transform": "translateX(1000px)", "-webkit-transform": "translateX(1000px)", "-ms-transform": "translateX(1000px)"});
−
An immunized person has antibodies against tetanus antigens - we bound spots of our purified tetanus antigens onto the specific surface of our chip. We then incubated the DiaChip with 33 µl of the blood serum for half an hour and measured the results.
+
    $('#second').css({"transform": "translateX(1000px)", "-webkit-transform": "translateX(1000px)", "-ms-transform": "translateX(1000px)"});
 +
    $('#third').css({"z-index": "100"});
 +
    $('#fourth').css({"z-index": "-100", "transform": "translateX(-2000px)", "-webkit-transform": "translateX(-2000px)", "-ms-transform": "translateX(-2000px)", "-webkit-transform": "translateX(-2000px)", "-ms-transform": "translateX(-2000px)"});
 +
    distance = 3;
 +
}
  
−
Take a look at the video and see for yourself how good our DiaChip performed.
+
else if (distance ==3) {
 +
console.log("Distance: "+distance);
 +
$('#first').css({"z-index": "-100", "transform": "translateX(-1000px)", "-webkit-transform": "translateX(-1000px)", "-ms-transform": "translateX(-1000px)", "-webkit-transform": "translateX(-1000px)", "-ms-transform": "translateX(-1000px)"});
 +
    $('#second').css({"transform": "translateX(2000px)", "-webkit-transform": "translateX(2000px)", "-ms-transform": "translateX(2000px)", "-webkit-transform": "translateX(2000px)", "-ms-transform": "translateX(2000px)"});
 +
    $('#third').css({"transform": "translateX(1000px)", "-webkit-transform": "translateX(1000px)", "-ms-transform": "translateX(1000px)"});
 +
    $('#fourth').css({"z-index": "100"});
 +
    distance = 4;
 +
}
  
−
And remember: that this isn’t an ELISA, we don’t need no expensive secondary Antibodies!
+
else if (distance ==4) {
−
</p>
+
console.log("Distance: "+distance);
−
</div>
+
$('#first').css({"z-index": "100"});
−
</div>
+
    $('#second').css({"z-index": "-100", "transform": "translateX(0)", "-webkit-transform": "translateX(0)", "-ms-transform": "translateX(0)"});
 +
    $('#third').css({"transform": "translateX(2000px)", "-webkit-transform": "translateX(2000px)", "-ms-transform": "translateX(2000px)"});
 +
    $('#fourth').css({"transform": "translateX(-1000px)", "-webkit-transform": "translateX(-1000px)", "-ms-transform": "translateX(-1000px)"});
 +
    distance = 1;
 +
}
  
 +
// see: http://stackoverflow.com/questions/1836105/how-to-wait-5-seconds-with-jquery //
 +
// wait until css-animation has finished. time is specified in css-transition property //
 +
setTimeout(function(){
 +
$("#buttonleft").bind("click", moveleft);
 +
}, 1000);
 +
}
  
  
−
<div id="agfp_results">
+
    $("#buttonright").bind("click", moveright);
−
<div class="floatbox right">
+
function moveright(e) {
−
<h2>
+
 
−
We measured the binding of anti-GFP to cell-free expressed GFP in blood serum
+
$("#buttonright").unbind();
−
</h2>
+
console.log("Click "+distance);
−
<p>
+
 
−
We expressed GFP with our own cell-free expression system and bound it to the surface of our DiaChip. We than ran some rabbit blood serum over the chip which we had spiked with anti-GFP antibodies. The results show that our cell-free proteins can be measured even in a complex solution such as blood serum. </p>
+
if (distance == 1) {
 +
$('#first').css({"z-index": "-100"});
 +
    $('#second').css({"transform": "translateX(2000px)", "-webkit-transform": "translateX(2000px)", "-ms-transform": "translateX(2000px)", "-webkit-transform": "translateX(2000px)", "-ms-transform": "translateX(2000px)"});
 +
    $('#third').css({"z-index": "100","transform": "translateX(1000px)", "-webkit-transform": "translateX(1000px)", "-ms-transform": "translateX(1000px)"});
 +
    $('#fourth').css({"transform": "translateX(-2000px)", "-webkit-transform": "translateX(-2000px)", "-ms-transform": "translateX(-2000px)"});
 +
    distance = 4;
 +
}
 +
 
 +
else if (distance ==2) {
 +
console.log("Distance: "+distance);
 +
$('#first').css({"transform": "translateX(-1000px)", "-webkit-transform": "translateX(-1000px)", "-ms-transform": "translateX(-1000px)"});
 +
    $('#second').css({"z-index": "-100"});
 +
    $('#third').css({"transform": "translateX(2000px)", "-webkit-transform": "translateX(2000px)", "-ms-transform": "translateX(2000px)", "-webkit-transform": "translateX(2000px)", "-ms-transform": "translateX(2000px)"});
 +
    $('#fourth').css({"z-index": "100","transform": "translateX(-1000px)", "-webkit-transform": "translateX(-1000px)", "-ms-transform": "translateX(-1000px)"});
 +
    distance = 1;
 +
}
 +
 
 +
else if (distance ==3) {
 +
console.log("Distance: "+distance);
 +
$('#first').css({"z-index": "100","transform": "translateX(0px)", "-webkit-transform": "translateX(0px)", "-ms-transform": "translateX(0px)"});
 +
    $('#second').css({"transform": "translateX(0)", "-webkit-transform": "translateX(0)", "-ms-transform": "translateX(0)"});
 +
    $('#third').css({"z-index": "-100"});
 +
    $('#fourth').css({"transform": "translateX(0)", "-webkit-transform": "translateX(0)", "-ms-transform": "translateX(0)"});
 +
    distance = 2;
 +
}
 +
 
 +
else if (distance ==4) {
 +
console.log("Distance: "+distance);
 +
$('#first').css({"transform": "translateX(1000px)", "-webkit-transform": "translateX(1000px)", "-ms-transform": "translateX(1000px)"});
 +
    $('#second').css({"z-index": "100","transform": "translateX(1000px)", "-webkit-transform": "translateX(1000px)", "-ms-transform": "translateX(1000px)"});
 +
    $('#third').css({"transform": "translateX(0)", "-webkit-transform": "translateX(0)", "-ms-transform": "translateX(0)"});
 +
    $('#fourth').css({"z-index": "-100"});
 +
    distance = 3;
 +
}
 +
 
 +
setTimeout(function(){
 +
$("#buttonright").bind("click", moveright);
 +
}, 1000);
 +
}
 +
});
 +
</script>
 +
 
 +
 
 +
 
 +
<div class="container">
 +
<div id="buttonright">
 +
<img src="https://static.igem.org/mediawiki/2015/b/b3/Freiburg_lightbulb_slider_30.png">
 
</div>
 
</div>
−
<div class="floatbox left">
+
<div id="buttonleft">
−
<div class="imagebox left">
+
<img src="https://static.igem.org/mediawiki/2015/4/4d/Freiburg_lightbulb_slider_27.png">
−
<img src="https://static.igem.org/mediawiki/2015/7/74/Freiburg_labjournal-roi_selection_20150817_429.png" width="100%"></img>
+
−
</div>
+
 
</div>
 
</div>
 +
  <div id="content-slider">
 +
      <div id="slider">  <!-- Slider container -->
 +
        <div id="mask">  <!-- Mask -->
 +
        <ul>
 +
        <li id="first" class="firstanimation">  <!-- ID for tooltip and class for animation -->
 +
        <div class="artboard">
 +
        <div class="sliderimage">
 +
        <img src="https://static.igem.org/mediawiki/2015/8/89/Freiburg_Slider-DIY_scaled.png" width="350px">
 +
        </div>
 +
        <div class="slidertext">
 +
        <h1>Building our own device</h1>
 +
        <p>The device orginally used, in collaboration with AG Roth, was a pricy machine based on rather simple physics. Therefore, we decided to build our own apparatus in a cost-efficient manner. We were able to produce reliable results with it and provided a construction plan. This plan will make it possible for future iGEM generations to built and use their own label-free protein array analysis tool.</p>
 +
        </div>
 +
        <div class="slidertext indent">
 +
        <p> Want to read <a href="https://2015.igem.org/Team:Freiburg/OwnDevice"> more?</a>
 +
        </p>
 +
        </div>
 +
        </div>
 +
        </li>
 +
        <li id="second" class="secondanimation">
 +
        <div class="artboard">
 +
        <div class="sliderimage">
 +
        <img src="https://static.igem.org/mediawiki/2015/b/b8/Freiburg_Slider-HumanPractice_scaled.png" width="350px">
 +
        </div>
 +
        <div class="slidertext">
 +
        <h1>Communicating science</h1>
 +
        <p>Diagnosing diseases fast and reliable has not just been an issue among medical staff, it has also been subject to public interest. This has lead us to ask for people's opinions regarding the DiaCHIP. Although the device is labeled as a product of synthetic biology, which has been problematic for the broad public according to a survey initiated by the Leopoldina (National academy of science), we recieved lot of positive feedback. </p>
 +
        </div>
 +
        <div class="slidertext indent">
 +
        <p> Want to read <a href="https://2015.igem.org/Team:Freiburg/Practices"> more?</a></p>
 +
        </div>
 +
        </div>
 +
        </li>
 +
        <li id="third" class="thirdanimation">
 +
        <div class="artboard">
 +
        <div class="sliderimage">
 +
        <img src="https://static.igem.org/mediawiki/2015/1/14/Freiburg_Slide-Modelling_scaled.png" width="350px">
 +
        </div>
 +
        <div class="slidertext">
 +
        <h1>Modeling cellfree expression</h1>
 +
        <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 and identified the factors limiting cell-free expression in the DiaCHIP. </p>
 +
        </div>
 +
        <div class="slidertext indent">
 +
        <p> Want to read <a href="https://2015.igem.org/Team:Freiburg/Modeling">more?</a> </p>
 +
        </div>
 +
        </div>
 +
        </li>
 +
        <li id="fourth" class="fourthanimation">
 +
        <div class="artboard">
 +
        <div class="sliderimage">
 +
        <img src="https://static.igem.org/mediawiki/2015/e/e4/Freiburg_Slider-Eigenblut.png" width="350px">
 +
        </div>
 +
        <div class="slidertext">
 +
        <h1>Measuring our own blood</h1>
 +
        <p>One of the most promising results came from the detection of anti-tetanus antibodies in human blood serum. The DiaCHIP analysis made it possible for us to distinguish serum samples from an individual before and after vaccination. Samples taken three weeks after vaccination produced positive signals, compared to negative results prior to antigen exposure. </p>
 +
        </div>
 +
        <div class="slidertext indent">
 +
        <p> Want to read <a href="https://2015.igem.org/Team:Freiburg/Results">more?</a></p>
 +
        </div>
 +
        </div>
 +
        </li>
 +
    </ul>
 +
        </div>  <!-- End Mask -->
 +
      </div>  <!-- End Slider Container -->
 +
  </div>
 
</div>
 
</div>
  
  
−
<div id="device_results">
 
−
<h1>
 
−
We built our very own, low-cost DiaChip device - and can measure with it!
 
−
</h1>
 
−
<div class="flexbox">
 
−
<img src="https://static.igem.org/mediawiki/2015/7/74/Freiburg_labjournal-roi_selection_20150817_429.png" width="80%"></img>
 
−
</div>
 
−
<p>
 
−
A green LED, two lenses and a regular SLR Camera is basically all you need to build your own iRIf device.
 
−
</p>
 
−
</div>
 
−
</div>
 
  
 
<div class="content_box">
 
<div class="content_box">
 +
<!-- Labjournal content goes in here -->
 +
  
−
<div class="flexbox">
+
<h1 class="sectionedit1">Project overview: The DiaCHIP</h1>
−
<a href="https://2015.igem.org/Team:Freiburg/Results/Assembling">
+
<div class="level1">
−
<img src="https://static.igem.org/mediawiki/2015/0/0f/Freiburg_homepage_chip_blood.png" width="100%">
+
<div class="image_box left">
−
</a>
+
<img align="left" alt="DiaCHIP_Sabi" src="https://static.igem.org/mediawiki/2015/a/af/Freiburg_DiaCHIP_Sabi.png" width="420px">
 +
</div>
 +
<p>
 +
<!--korrigiert von Philipp05/09/15-->
 +
<!--neu von Ramona08/09/15-->
 +
The DiaCHIP is an innovative tool to screen for a broad range of antibodies present in serum. Antibodies can be an indicator for  an immune response towards an infection or a succesfull vaccination. Antibodies also play a role in the diagnosis of autoimmune diseases. Especially the ability to differentiate between life threatening diseases and a mild infection within minutes bears the potential to save lifes. 
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Spotting diseases by detecting correspondent antibodies in a patient's serum is an established method in  <a class="wikilink1" href="https://2015.igem.org/Team:Freiburg/Diagnostics" title="diagnostics_today">modern diagnostics</a>. The DiaCHIP makes this possible using a single blood sample.
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The key feature of the DiaCHIP concept is the combination of on-demand protein synthesis and a novel method of label-free detection packed in 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.  (LINK – new device).
  
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<p> Click on one of the images to get further insight how we build up our DiaCHIP </p>
 
  
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<b>Step 1: Preparing the DiaCHIP by protein synthesis</b>
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Pior to screening for antibody-antigen interactions, the antigens have to be synthesized and immobilized in a microarray set-up. Facilitated by a copy mechanism, which 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 redudant.
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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 PDMS 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 this other surface by diffusion. Complex chemistry ensures that target proteins are specifically immobilized on this surface, while components of the expression mix can be washed away before sample analysis.
  
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<b>Step 2: Measuring serum samples by iRIf</b>
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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 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 thin layers, binding events can be recorded in real-time.
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After weeks of opimizing the different components of the DiaCHIP, we reveal our great results. The highlight of our project was reached with the successful <a href="https://2015.igem.org/Team:Freiburg/Results">detection of antibodies in our own blood!</a>
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<a class="wikilink1" href="/igem2015/doku.php?id=tag:info&amp;do=showtag&amp;tag=info" rel="tag" title="tag:info">info</a>
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Revision as of 23:10, 9 September 2015

""

  • Building our own device

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

    Want to read more?

  • Communicating science

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

    Want to read more?

  • Modeling cellfree 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 and identified the factors limiting cell-free expression in the DiaCHIP.

    Want to read more?

  • Measuring our own blood

    One of the most promising results came from the detection of anti-tetanus antibodies in human blood serum. The DiaCHIP analysis made it possible for us to distinguish serum samples from an individual before and after vaccination. Samples taken three weeks after vaccination produced positive signals, compared to negative results prior to antigen exposure.

    Want to read more?

Project overview: The DiaCHIP

DiaCHIP_Sabi

The DiaCHIP is an innovative tool to screen for a broad range of antibodies present in serum. Antibodies can be an indicator for an immune response towards an infection or a succesfull vaccination. Antibodies also play a role in the diagnosis of autoimmune diseases. Especially the ability to differentiate between life threatening diseases and a mild infection within minutes bears the potential to save lifes.
Spotting diseases by detecting correspondent antibodies in a patient's serum is an established method in modern diagnostics. The DiaCHIP makes this possible using a single blood sample.
The key feature of the DiaCHIP concept is the combination of on-demand protein synthesis and a novel method of label-free detection packed in 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. (LINK – new device).

Step 1: Preparing the DiaCHIP by protein synthesis
Pior to screening for antibody-antigen interactions, the antigens have to be synthesized and immobilized in a microarray set-up. Facilitated by a copy mechanism, which 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 redudant.
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 PDMS 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. The template slide is placed in close proximity to the future protein array enabling the expressed proteins to reach this other surface by diffusion. Complex chemistry ensures that target proteins are specifically immobilized on this surface, while components of the expression mix can be washed away before sample analysis.

Step 2: Measuring serum samples by 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 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 thin layers, binding events can be recorded in real-time.

After weeks of opimizing the different components of the DiaCHIP, we reveal our great results. The highlight of our project was reached with the successful detection of antibodies in our own blood!