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. 

Figure. Similarity between the Bridge-It^® technology and the PINCER^TM technology. The PINCER (binder) 1 and 2 can be either a DNA apatmer, a RNA aptamer, an antibody, a peptide or a ligand.

Comparison between Pincer^TM assay and ELISA (hCRP)

Comparison of the hCRP serum spiked samples measured by ELISA and Pincer^TM assay

The Pincer^TM products have been developed in part with support from the National Institutes of Health, through STTR Phase I and Phase II Grants, Award No.: GM079891. Patent pending.