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Brief History of Our Bridge-It® and PINCER® Technologies

Mediomics’ Bridge-it® assay (Fig. 1A) was based on a simple idea of splitting the DNA binding site for a protein into two DNA “half-sites”. Each of the resulting “half-sites” contains a short complementary single-stranded region of the length designed to introduce some propensity for the two DNA “half-sites” to associate recreating the duplex containing the fully functional protein binding site. This propensity is designed to be low such that in the absence of the protein only a small fraction of DNA half-sites will associate. Fig. 1B illustrates how the Bridge-It® technology was evolved into the PINCER® technology. Instead of splitting the DNA duplex containing the natural binding site for a protein into the two “half-sites”, two PINCERs (binders) recognizing two different epitopes of the target are used as functional equivalents of the “half-sites”. Short signaling molecules containing the matching pair of fluorophores are attached to the two PINCERs via a flexible linker (Fig. 1B). In the absence of the target, the two PINCERs (half-sites) will not associate. Upon addition of the target, the preferential binding of the target will drive the association of the two PINCERs (half-sites) resulting in fluorescence signal change.