Systematic study of steric and spatial contributions to molecular recognition by non-covalent imprinted polymers.

Although molecular imprinting is a widely accepted method for producing template specific polymers, the general rules for prediction and control of the binding and catalytic properties of these materials are still not fully understood. One reason for this is the problematic structural analysis of the active sites in the polymers, which are not amenable to X-ray crystallography or microscopic techniques due to their amorphous and heterogeneous nature. Therefore, molecular probes have been the most informative agents for the analysis of the structure of active sites. This paper focuses on the steric and geometrical aspects of shape recognition in non-covalent imprinted polymers, with particular effort to minimize other factors contributing to molecular recognition by the polymers. Chiral amine compounds with systematic changes in spatial, distal and conformational components of sterically controlled molecular recognition were investigated for use as non-covalent imprinted polymers. Chromatographic studies revealed that steric and spatial interactions influence the selectivity properties of imprinted polymers in a predictable fashion.

Development of an aspartic acid-based cross-linking monomer for improved bioseparations.

Improved specificity and binding affinity by molecularly imprinted polymers is possible by development of novel functional materials. Furthermore, increasing the cross-link density of imprinted polymers by using cross-linking functional groups was anticipated to improve polymer molecular recognition. A novel cross-linking monomer derived from an L-aspartic acid precursor was synthesized and employed in molecularly imprinted polymers to mimic more closely the scaffolding of proteins, and thus provide more protein-like selectivity. Chromatographic results revealed a more than 7-fold improvement in polymers imprinted using the new monomer versus a traditionally formulated polymer imprinted with methacrylic acid as the functional monomer.

A comparison of flexible and constrained haptens in eliciting antibody catalysts for paraoxon hydrolysis.

A new amine-oxide hapten was employed as an antigen, producing seven monoclonal antibodies (mAbs) from a panel of 20 that catalyzed paraoxon hydrolysis. The current hapten design differs from that previously described in that the molecule is inherently more flexible than its constrained predecessor. One of the seven antibody catalysts, mAb 1H9, showed the highest activity and was selected for detailed study. At pH = 8.77, the catalytic hydrolysis of paraoxon by mAb 1H9 followed Michaelis Menten kinetics affording a k(cat) = 3.73 x 10(-4) min(-1) and a Km = 1.12 mM with a rate acceleration k(cat)/k(uncat) = 56. The hapten was found to be a competitive inhibitor of antibody-catalyzed paraoxon hydrolysis with a Ki = 0.54 mM. A comparison of both the number and proficiency of antibody catalysts obtained when utilizing a flexible versus constrained hapten indicates that, for paraoxon hydrolysis, constrained haptens elicit superior catalysts, suggesting that further development should begin with the use of constrained haptens in producing more proficient antibody catalysts for paraoxon hydrolysis.