Furthermore, the binding of biomarker with aptamer of Kd of nM levels can usually break the hybridization with Tm of 50 degree celsius. The binding with Kd of micro M can usually break the hybridization with Tm of less than 40 degree celsius.
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We followed established protocols for ELONA, an ELISA assay modified to accommodate aptamers, in order to determine the dissociation constant. We used a protocol modified from Sang et al to determine the Kd value using the Hill equation, a Scatchard plot, and non-linear regression. However, the results were inconclusive so we decided to find our Kd value through the 3D modeling software, UNAFold. <br/><br/>
As our results show, the resulting aptamer probes were successful and produced satisfactory results that lined up with our mathematical predictions, thus validating our design guidelines (please refer to our results page).<br/><br/>
The results demonstrated that the aptamer probes produced satisfactory results and aligned with our mathematical predictions, thus validating our design guidelines. Please refer to our results page for further details.<br/><br/>
Lastly, we have devised a simple workflow for the detection and quantification of our biomarkers of interest using the Hill equation. For more information, please see our Modeling page.
