Molecularly imprinted hydrogel displaying reduced non-specific binding and improved protein recognition.

A novel approach for enhancing protein recognition in molecularly imprinted hydrogel (MIH) is presented. This approach was developed based on the hypothesis that the number of specific binding sites created in the previously described MIH is very small, thus attempts to enhance the capacity result in most cases in additional non-specific binding and loss of selectivity. Thus, blocking the non-specific binding sites could lead to higher capacities and better selectivity. To test this hypothesis, MIH interpenetrating networks designed to block non-specific binding sites were synthesized using two separate stages of polymerization. Re-binding of the template protein (lysozyme) and a competitor protein (cytochrome C) was measured, and the results were compared with the similar experiment performed using a control non-imprinted hydrogel and a "conventional" MIH. The imprinting efficacy of the MIH interpenetrating network was found to be much higher than that of the controls. Furthermore, competitive adsorption assays have demonstrated the superiority of the new formulation.