Team:Potsdam Bioware

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<h2>Direct Antibody Identification, Maturation and Production in CHO-Cells</h2>
 
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== Antibody Generation System - Maturation, Selection and Production in CHO Cells ==
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=== Project Description ===
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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.
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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.  
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<h3>Team<h3>
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      <div class="team_box">Mario Köhler</div>
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      <div class="team_box">Tom Scheidt</div>
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      <div class="team_box">Christopher Kressler</div>
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      <div class="team_box">Tobias Priemel</div>
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      <div class="team_box">Maria Kunert</div>
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      <div class="team_box">Sascha Lehnort</div>
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      <div class="team_box">Norman Hein</div>
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<font size="-4"; color="grey">primary contact: Kristian Müller<br>
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<a href="http://www.syntbio.net"> http://www.syntbio.net</a> <a href="mailto:kristian@syntbio.net">kristian@syntbio.net</a></font></center></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

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