Team:Potsdam Bioware

From 2012.igem.org

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<h2>Direct Antibody Identification, Maturation and Production in CHO-Cells</h2>
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<h2>Antibody Generation System - Maturation, Selection and Production in CHO Cells</h2>
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<h3>Project Description</h3>
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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. </div>
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Revision as of 16:52, 15 September 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.

Team


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