Student award for outstanding research winner in the Ph.D. category for the 2017 society for biomaterials annual meeting and exposition, april 5–8, 2017, Minneapolis, Minnesota: Characterization of protein interactions with molecularly imprinted hydrogels that possess engineered affinity for high isoelectric point biomarkers

Abstract

Molecularly imprinted polymers (MIPs) with selective affinity for protein biomarkers could find extensive utility as environmentally robust, cost-efficient biomaterials for diagnostic and therapeutic applications. In order to develop recognitive, synthetic biomaterials for prohibitively expensive protein biomarkers, we have developed a molecular imprinting technique that utilizes structurally similar, analogue proteins. Hydrogel microparticles synthesized by molecular imprinting with trypsin, lysozyme, and cytochrome c possessed an increased affinity for alternate high isoelectric point biomarkers both in isolation and plasma-mimicking adsorption conditions. Imprinted and non-imprinted P(MAA-co-AAm-co-DEAEMA) microgels containing PMAO-PEGMA functionalized polycaprolactone nanoparticles were net-anionic, polydisperse, and irregularly shaped. MIPs and control non-imprinted polymers (NIPs) exhibited regions of Freundlich and BET isotherm adsorption behavior in a range of non-competitive protein solutions, where MIPs exhibited enhanced adsorption capacity in the Freundlich isotherm regions. In a competitive condition, imprinting with analogue templates (trypsin, lysozyme) increased the adsorption capacity of microgels for cytochrome c by 162% and 219%, respectively, as compared to a 122% increase provided by traditional bulk imprinting with cytochrome c. Our results suggest that molecular imprinting with analogue protein templates is a viable synthetic strategy for enhancing hydrogel-biomarker affinity and promoting specific protein adsorption behavior in biological fluids.

Label-Free Detection of Tear Biomarkers Using Hydrogel-Coated Gold Nanoshells in a Localized Surface Plasmon Resonance-Based Biosensor

“…12 While the potential of protein imprinted polymers is alluring and remains a continued topic of research, there has been minimal success in achieving truly selective polymers, largely due to challenges associated with protein size and structural complexity. 13–15…”

QCM‐D assay for quantifying the swelling, biodegradation, and protein adsorption of intelligent nanogels

“…For the purpose of this study, lysozyme was selected as a model ligand. We knew from previous studies that lysozyme binds in significant quantity to poly(methacrylic acid‐ co ‐acrylamide) copolymers in aqueous buffer . We infused lysozyme dissolved in PBS at up to 5 mg/mL over the nanogel‐coated and uncoated sensors.…”

Synthetic networks with tunable responsiveness, biodegradation, and molecular recognition for precision medicine applications

“…The extent of nanogel functionalization with peptides or proteins was quantified using a Micro BCA colorimetric assay (Thermo Fisher Scientific), as described previously ( 39 ). Modified or unmodified nanogels, suspended at 2 mg/ml in 1× PBS (pH 7.4 ± 0.05), were combined at an equal volume ratio with Micro BCA working reagent and mixed for 2 hours at 37°C (constant mixing).…”