Applying the sample taken three weeks after vaccination to the DiaCHIP resulted in a significant signal at the antigen spot, whereas no signal was obtained with the sample assumed to be negative. But convince yourself of how great the DiaCHIP performed. The video on the right shows these measurements right next to each other in real-time.
Besides tetanus, some other antigens of immunological relevance were taken into account. See all the results we obtained in terms of diagnostics, including an approach for quantification of the measurements.
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Figure 2B shows the binding of anti-GFP to GFP as it was established before, measured with our own device. | Figure 2B shows the binding of anti-GFP to GFP as it was established before, measured with our own device. | ||
Look here, to see how you can <a href="https://2015.igem.org/Team:Freiburg/Results/Own_Device">build your own device</a> for label-free detection of antibody binding on a protein microarray needing two lenses, a camera and a little bit of fine feeling (??) sensitivety/sensitiveness/finesse/sure instincts. | Look here, to see how you can <a href="https://2015.igem.org/Team:Freiburg/Results/Own_Device">build your own device</a> for label-free detection of antibody binding on a protein microarray needing two lenses, a camera and a little bit of fine feeling (??) sensitivety/sensitiveness/finesse/sure instincts. | ||
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Revision as of 17:55, 12 September 2015
![](https://static.igem.org/mediawiki/2015/8/85/Freiburg_wiki_background_2_parted_left.png)
![](https://static.igem.org/mediawiki/2015/f/f4/Freiburg_wiki_background_2_parted_right.png)
Nicole: Hab ich es richtig verstanden, dass hier nur eure Highlights and Ergebnissen dargestellt werden sollen (mit verlinkung zu den anderen Ergebnissen)? Was haltet ihr von einer kurzen Einleitung... In the last months we (aimed to) developed a diagnostic tool which enables [...] We succeded to detect anti-tetanus antibodies in human serum, develop/generate our own cell-free mix...
mehr Formulierungen wie: we demonstrated..., we (successfully) tested, we achieved...
Detection of anti-Tetanus antibodies in human blood serum
![](https://static.igem.org/mediawiki/2015/7/74/Freiburg_labjournal-roi_selection_20150817_429.png)
We specifically detected anti-Tetanus antibodies in human blood serum. To test the DiaCHIP under real-life conditions, we analyzed the blood serum of a vaccinated person for the presence of anti-Tetanus antibodies. We compared blood samples before and after vaccination and could directly detect its effect. To capture the antibodies, the corresponding antigen was expressed in E. coli, purified by His-tag affinity purification and spotted on a specific Ni-NTA surface.
Detection of anti-GFP antibodies in rabbit serum using cell-free expressed GFP
![](https://static.igem.org/mediawiki/2015/7/74/Freiburg_labjournal-roi_selection_20150817_429.png)
To show that cell-free expressed proteins can be immobilized on the protein surface while maintaining their antibody binding properties, the expression mix was spotted on a specific Ni-NTA surface after GFP expression. Cell-free expressed GFP was spotted on a specific Ni-NTA slide and flushed with serum of a rabbit immunized against GFP. The lowest spot consisting of cell-free expressed GFP shows a significant signal compared to the negative control (middle spot).
Building our very own, low-cost DiaCHIP measuring device.
The measuring device we used in collaboration with the AG Roth (ZBSA) to detect antibody binding events is a really expensive machine, but the physics which it is based on is rather simple. Our self-built device consistent of not much more than two lenses and a camera (A) reliably detected the binding of anti-GFP to GFP (B). To enable future iGEM teams to profit from this label-free detection method as we did, a cost-efficient and easy to rebuild variant of the device was developed (figure 2A).
Specific surfaces
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Own cell-free mix
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