Team:Potsdam Bioware

From 2012.igem.org

(Difference between revisions)
(→Project Description)
 
(81 intermediate revisions not shown)
Line 1: Line 1:
-
==Direct Antibody Identification, Maturation and Production in CHO-Cells==
+
{{:Team:Potsdam_Bioware/header}}
-
<div class="box_round">
+
<div class="box_round gradient_grey">
-
===Project Description===
+
== Antibody Generation System - Maturation, Selection and Production in CHO Cells ==
-
<hr/>
+
=== Project Description ===
-
Antibodies are versatile molecules for research and therapy and they are currently revolutionizing the market of biopharmaceuticals. To date, antibodies are generated in a laborious and time consuming manner. Either animal immunization followed by hybridoma technology or phage display is used to identify desired genes, which then need to be transferred to a production cell line such as CHO. We designed a streamlined workflow that incorporates all steps of antibody generation in CHO cells. Our plan is to stably transfect an antibody-fragment library in CHO cells. Antibody maturation is mimicked by further diversifying this library using the enzyme Activation-Induced cytidine Deaminase (AID), which is known to induce somatic hypermutation. Next, we plan to test and deploy a versatile and continuous viral selection system for clones producing the desired antibodies. Finally, we will try to employ a genetic switch to go from surface expression to soluble production of antibodies.
+
Antibodies are of utmost importance for research and therapy but their generation is laborious and time consuming. We established a novel streamlined workflow for obtaining antibodies by incorporating all natural steps such as antibody maturation, selection and production in one genetic system implemented into a eukaryotic cell line. We stably transfect an antibody construct into CHO cells and mimic maturation by using the enzyme AID (activation-induced deaminase), which is known to induce somatic hypermutation. For selection, we are testing and deploying a versatile and continuous viral system as well as magnetic beads and cell sorting. Finally, a genetic switch enables the transition from surface expression to production of soluble antibodies. In addition, we pursue phage display with an antibody fragment to study mutation rate and evolution by AID in prokaryotes. Our system supersedes animal immunization and the smooth process will increase the ready availability of antibodies in various formats.  
</div>
</div>
 +
<html>
-
<html><center>
+
-
<div id="clustrmaps-widget"></div>
+
<div class="rollover">
-
<script type="text/javascript">
+
  <div id="first_row">
-
var _clustrmaps = {'url' : https://2012.igem.org/Team:Potsdam_Bioware', 'user' : 1035150, 'server' : '4', 'id' : 'clustrmaps-widget', 'version' : 1, 'date' : '2012-08-08', 'lang' : 'en', 'corners' : 'square' };(function (){ var s = ocument.createElement('script'); s.type = 'text/javascript'; s.async = true; s.src = http://www4.clustrmaps.com/counter/map.js'; var x = document.getElementsByTagName('script')[0]; x.parentNode.insertBefore(s, x);})();
+
  <a id="rollover-image" href="https://2012.igem.org/Team:Potsdam_Bioware/Project/At_a_Glance"></a>
-
</script>
+
  <a id="rollover-image-2" href="https://2012.igem.org/Team:Potsdam_Bioware/Project/Part_AID"></a>
-
<noscript>
+
  <a id="rollover-image-3" href="https://2012.igem.org/Team:Potsdam_Bioware/Project/Part_Antibody"></a>
-
<ahref="http://www4.clustrmaps.com/user/3d4fcb8e"><img src="http://www4.clustrmaps.com/stats/maps-no_clusters/2012.igem.org-Team-Potsdam_Bioware-thumb.jpg" alt="Locations of visitors to this page" /></a>
+
  <a id="rollover-image-8" href="https://2012.igem.org/Team:Potsdam_Bioware/Project/Part_Virus"></a>
-
</noscript>
+
  </div>
-
</center></html>
+
  <div id="second_row">
 +
  <a id="rollover-image-7" href="https://2012.igem.org/Team:Potsdam_Bioware/Biobricks/Overview"></a>
 +
  <a id="rollover-image-6" href="https://2012.igem.org/Team:Potsdam_Bioware/Project/Potsdam%20Standard"></a>
 +
  <a id="rollover-image-5" href="https://2012.igem.org/Team:Potsdam_Bioware/SocialBrick"></a>
 +
  <a id="rollover-image-4" href="https://2012.igem.org/Team:Potsdam_Bioware/Project/Modeling"></a>
 +
  </div>
 +
</div>
-
<!-- *** What falls between these lines is the Alert Box!  You can remove it from your pages once you have read and understood the alert ***
 
-
 
-
<html>
 
-
<div id="box" style="width: 700px; margin-left: 137px; padding: 5px; border: 3px solid #000; background-color: #fe2b33;">
 
-
<div id="template" style="text-align: center; font-weight: bold; font-size: large; color: #f6f6f6; padding: 5px;">
 
-
This is a template page. READ THESE INSTRUCTIONS.
 
-
</div>
 
-
<div id="instructions" style="text-align: center; font-weight: normal; font-size: small; color: #f6f6f6; padding: 5px;">
 
-
You are provided with this team page template with which to start the iGEM season.  You may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki.  You can find some examples <a href="https://2009.igem.org/Help:Template/Examples">HERE</a>.
 
-
</div>
 
-
<div id="warning" style="text-align: center; font-weight: bold; font-size: small; color: #f6f6f6; padding: 5px;">
 
-
You <strong>MUST</strong> have all of the pages listed in the menu below with the names specified. PLEASE keep all of your pages within your teams namespace. 
 
-
</div>
 
-
</div>
 
</html>
</html>
-
*** End of the alert box ***
 
-
*<b>"Go, Krauts, go"</b> - Best german team 2012 is Potsdam (same like last year) :)!
 
-
 
+
<div class="box_round no_padding">
-
{|align="justify"
+
{{:Team:Potsdam_Bioware/slider}}
-
|You can write a background of your team here.  Give us a background of your team, the members, etc.  Or tell us more about something of your choosing.
+
</div>
-
|[[Image:Potsdam_Bioware_logo.png|200px|right|frame]]
+
-
|-
+
-
|
+
-
''Tell us more about your project.  Give us background.  Use this as the abstract of your project.  Be descriptive but concise (1-2 paragraphs)''
+
-
|[[Image:Potsdam_Bioware_team.png|right|frame|Your team picture]]
+
-
|-
+
-
|
+
-
|align="center"|[[Team:Potsdam_Bioware | Team Potsdam_Bioware]]
+
-
|}
+
-
-->
+
-
<!--- The Mission, Experiments --->
+
-
 
+
-
{| style="color:#1b2c8a;background-color:#0c6;" border="1" bordercolor="#fff" width="62%" align="center"
+
-
!align="center"|[[Team:Potsdam_Bioware|Home]]
+
-
!align="center"|[[Team:Potsdam_Bioware/Team|Team]]
+
-
!align="center"|[https://igem.org/Team.cgi?year=2012&team_name=Potsdam_Bioware Official Team Profile]
+
-
!align="center"|[[Team:Potsdam_Bioware/Project|Project]]
+
-
!align="center"|[[Team:Potsdam_Bioware/Parts|Parts Submitted to the Registry]]
+
-
!align="center"|[[Team:Potsdam_Bioware/Modeling|Modeling]]
+
-
!align="center"|[[Team:Potsdam_Bioware/Notebook|Notebook]]
+
-
!align="center"|[[Team:Potsdam_Bioware/Safety|Safety]]
+
-
!align="center"|[[Team:Potsdam_Bioware/Attributions|Attributions]]
+
-
|}
+
<html>
<html>
-
<head>
+
<center>
-
<style type="text/css">
+
<div id="clustrmaps-widget">
 +
</div><script type="text/javascript">var _clustrmaps = {'url' : 'https://2012.igem.org/Team:Potsdam_Bioware', 'user' : 1035150, 'server' : '4', 'id' : 'clustrmaps-widget', 'version' : 1, 'date' : '2012-09-22', 'lang' : 'en', 'corners' : 'square' };(function (){ var s = document.createElement('script'); s.type = 'text/javascript'; s.async = true; s.src = 'http://www4.clustrmaps.com/counter/map.js'; var x = document.getElementsByTagName('script')[0]; x.parentNode.insertBefore(s, x);})();</script><noscript><a href="http://www4.clustrmaps.com/user/3d4fcb8e"><img src="http://www4.clustrmaps.com/stats/maps-no_clusters/2012.igem.org-Team-Potsdam_Bioware-thumb.jpg" alt="Locations of visitors to this page" /></a></noscript>
-
body {
+
<font size="-4"; color="grey">primary contact: Kristian Müller<br>
-
+
<a href="http://www.syntbio.net"> http://www.syntbio.net</a> <a href="mailto:kristian@syntbio.net">kristian@syntbio.net</a></font></center></html>
-
/* IE10 Consumer Preview */
+
-
background-image: -ms-linear-gradient(left, #FFFFFF 0%, #999999 100%);
+
-
 
+
-
/* Mozilla Firefox */
+
-
background-image: -moz-linear-gradient(left, #FFFFFF 0%, #999999 100%);
+
-
 
+
-
/* Opera */
+
-
background-image: -o-linear-gradient(left, #FFFFFF 0%, #999999 100%);
+
-
 
+
-
/* Webkit (Safari/Chrome 10) */
+
-
background-image: -webkit-gradient(linear, left top, right top, color-stop(0, #FFFFFF), color-stop(1, #999999));
+
-
 
+
-
/* Webkit (Chrome 11+) */  
+
-
background-image: -webkit-linear-gradient(left, #FFFFFF 0%, #999999 100%);
+
-
 
+
-
/* W3C Markup, IE10 Release Preview */
+
-
background-image: linear-gradient(to right, #FFFFFF 0%, #999999 100%);
+
-
}
+
-
 
+
-
.box_round {
+
-
  padding: 5px;
+
-
  border-radius: 15px;
+
-
  background: #cb9840;
+
-
  background: -moz-linear-gradient(top,  #cb9840 0%, #a77b34 100%);
+
-
  background: -webkit-gradient(linear, left top, left bottom, color-stop(0%,#cb9840), color-stop(100%,#a77b34));
+
-
  background: -webkit-linear-gradient(top,  #cb9840 0%,#a77b34 100%);
+
-
  background: -o-linear-gradient(top,  #cb9840 0%,#a77b34 100%);
+
-
  background: -ms-linear-gradient(top,  #cb9840 0%,#a77b34 100%);
+
-
  background: linear-gradient(to bottom,  #cb9840 0%,#a77b34 100%);
+
-
  filter: progid:DXImageTransform.Microsoft.gradient( startColorstr='#cb9840', endColorstr='#a77b34',GradientType=0 );
+
-
  box-shadow:8px 8px 8px #666;
+
-
}
+
-
 
+
-
</style>
+
-
</head>
+
-
</html>
+

Latest revision as of 13:32, 16 November 2012


Antibody Generation System - Maturation, Selection and Production in CHO Cells

Project Description

Antibodies are of utmost importance for research and therapy but their generation is laborious and time consuming. We established a novel streamlined workflow for obtaining antibodies by incorporating all natural steps such as antibody maturation, selection and production in one genetic system implemented into a eukaryotic cell line. We stably transfect an antibody construct into CHO cells and mimic maturation by using the enzyme AID (activation-induced deaminase), which is known to induce somatic hypermutation. For selection, we are testing and deploying a versatile and continuous viral system as well as magnetic beads and cell sorting. Finally, a genetic switch enables the transition from surface expression to production of soluble antibodies. In addition, we pursue phage display with an antibody fragment to study mutation rate and evolution by AID in prokaryotes. Our system supersedes animal immunization and the smooth process will increase the ready availability of antibodies in various formats.


Antibody module: In the first step, an antibody gene is integrated into the genome of CHO cells.

Antibody module: The CHO cell expresses the antibody on the cell surface.

Mutation module: The enzyme AID mutates the antibody gene -> maturation

Selection Module: A virus infects cells with high affine antibodies. These cells survive.

The selected cells are switched to antibody secreting cells with Cre-recombinase.

primary contact: Kristian Müller
http://www.syntbio.net kristian@syntbio.net

Retrieved from "http://2012.igem.org/Team:Potsdam_Bioware"