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Team:WPI-Worcester/Proof-of-Principle
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<p><h9>Agglutination Assay</h9></p><p>We were able to successfully achieve the results we wanted with the Agglutination Assay. When antibodies are in the presence of their matching antigen pairs, they will bind to each other. Antibodies have two antigen binding sites which allows a network of antibody-antigen expressing bacteria to form. This network manifests on a large scale as a mat of bacteria on the bottom of the containing vessel, as opposed to a solid dot formed by unagglutinated antibodies. We based our assay off of <a href="http://www.plosntds.org/article/info%3Adoi%2F10.1371%2Fjournal.pntd.0001946">this paper</a>.</p> | <p><h9>Agglutination Assay</h9></p><p>We were able to successfully achieve the results we wanted with the Agglutination Assay. When antibodies are in the presence of their matching antigen pairs, they will bind to each other. Antibodies have two antigen binding sites which allows a network of antibody-antigen expressing bacteria to form. This network manifests on a large scale as a mat of bacteria on the bottom of the containing vessel, as opposed to a solid dot formed by unagglutinated antibodies. We based our assay off of <a href="http://www.plosntds.org/article/info%3Adoi%2F10.1371%2Fjournal.pntd.0001946">this paper</a>.</p> | ||
| - | <p>For our proof of principle agglutination, we used surface-bound YFP(Yellow | + | <p>For our proof of principle agglutination, we used surface-bound YFP(Yellow Fluorescent Protein) and a corresponding GFP(Green Fluorescent Protein)/YFP antibody (the part of the protein that the antibody binds to is not affected by the part that causes the color change.) We used both of the surface expression proteins we had at our disposal: BclA (the one we made) and INP (a pre-existing biobrick). As controls, we used internally expressed YFP as well as a protein that was externally expressed but did not match with the antibody (we used the CAEV protein we made).</p> |
<p><center><img src="https://static.igem.org/mediawiki/2014/7/75/WPI_AgglutinationResults.png"/></center></p> | <p><center><img src="https://static.igem.org/mediawiki/2014/7/75/WPI_AgglutinationResults.png"/></center></p> | ||
Revision as of 15:07, 17 October 2014
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Team:WPI-Worcester
From 2014.igem.org
Proof of Principle
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We were able to successfully achieve the results we wanted with the Agglutination Assay. When antibodies are in the presence of their matching antigen pairs, they will bind to each other. Antibodies have two antigen binding sites which allows a network of antibody-antigen expressing bacteria to form. This network manifests on a large scale as a mat of bacteria on the bottom of the containing vessel, as opposed to a solid dot formed by unagglutinated antibodies. We based our assay off of this paper.
For our proof of principle agglutination, we used surface-bound YFP(Yellow Fluorescent Protein) and a corresponding GFP(Green Fluorescent Protein)/YFP antibody (the part of the protein that the antibody binds to is not affected by the part that causes the color change.) We used both of the surface expression proteins we had at our disposal: BclA (the one we made) and INP (a pre-existing biobrick). As controls, we used internally expressed YFP as well as a protein that was externally expressed but did not match with the antibody (we used the CAEV protein we made).
This photo clearly shows that the wells containing the bacteria with the matching antigen on the surface formed agglutinated mats while the control wells did not agglutinate and formed dots at the bottom of the wells.
