Team:Freiburg/Description

From 2011.igem.org

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(Plastic binding domain)
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====Plastic binding domain====
====Plastic binding domain====
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One oft the issues of our project „lab in a cell“ was to use endogenous proteins produced by the cell itself for specific purification and hereby to replace expensive columns. The „precipitator“ designed by our team contains a protein binding domain which complexes nickel and thus enables the binding of His-tagged proteins. After cell lysis the “precipitator” is freely dissolved in the cell lysate. To be able to insolate the “precipitator”-His-tagged-protein-complex from the other cell components it has to be immobilized by another protein domain.  
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One oft the issues of our project „lab in a cell“ was to use endogenous proteins produced by the cell itself for specific purification and hereby to replace expensive columns. The „precipitator“ designed by our team contains a protein binding domain which complexes nickel and thus enables the binding of His-tagged proteins. After cell lysis the “precipitator” is freely dissolved in the cell lysate. To be able to isolate the “precipitator”-His-tagged-protein-complex from the other cell components it has to be immobilized by another protein domain.  
During routine phage display of random peptide libraries, phages were found that bound directly to the plastic surface of the used plastic micro titer plates. The number of plastic binding phages obtained during the phage display experiments depended on the saturation of the plastic micro titer plates with target protein for the antibody-binding phages and could be reduced by the use of blocking proteins as BSA or non-fat milk. Plastic binding phages were resistant to washing steps with PBS alone as well as to PBS in combination with BSA or non-fat dry milk. It was shown that plastic binding phages were even more difficult to recover by acid elution than the “normal” antibody binding phages (Adey et al., 1995).  
During routine phage display of random peptide libraries, phages were found that bound directly to the plastic surface of the used plastic micro titer plates. The number of plastic binding phages obtained during the phage display experiments depended on the saturation of the plastic micro titer plates with target protein for the antibody-binding phages and could be reduced by the use of blocking proteins as BSA or non-fat milk. Plastic binding phages were resistant to washing steps with PBS alone as well as to PBS in combination with BSA or non-fat dry milk. It was shown that plastic binding phages were even more difficult to recover by acid elution than the “normal” antibody binding phages (Adey et al., 1995).  

Revision as of 16:03, 18 September 2011


This is the wiki page
of the Freiburger student
team competing for iGEM 2011.
Thank you for your interest!