Computationally Designed Epitope-Mediated Imprinted Polymers versus Conventional Epitope Imprints for the Detection of Human Adenovirus in Water and Human Serum Samples.

Detection of pathogenic viruses for point-of-care applications has attracted great attention since the COVID-19 pandemic. Current virus diagnostic tools are laborious and expensive, while requiring medically trained staff. Although user-friendly and cost-effective biosensors are utilized for virus detection, many of them rely on recognition elements that suffer major drawbacks. Herein, computationally designed epitope-imprinted polymers (eIPs) are conjugated with a portable piezoelectric sensing platform to establish a sensitive and robust biosensor for the human pathogenic adenovirus (HAdV). The template epitope is selected from the knob part of the HAdV capsid, ensuring surface accessibility. Computational simulations are performed to evaluate the conformational stability of the selected epitope. Further, molecular dynamics simulations are executed to investigate the interactions between the epitope and the different functional monomers for the smart design of eIPs. The HAdV epitope is imprinted via the solid-phase synthesis method to produce eIPs using in silico-selected ingredients. The synthetic receptors show a remarkable detection sensitivity (LOD: 102 pfu mL-1) and affinity (dissociation constant (Kd): 6.48 × 10-12 M) for HAdV. Moreover, the computational eIPs lead to around twofold improved binding behavior than the eIPs synthesized with a well-established conventional recipe. The proposed computational strategy holds enormous potential for the intelligent design of ultrasensitive imprinted polymer binders.

Cancer biomarker detection in human serum samples using nanoparticle decorated epitope-mediated hybrid MIP.

The determination of disease-associated molecules at trace amounts is a key factor for early and efficient diagnosis from human body fluids. Herein, an ultrasensitive electrochemical sensor based on hybrid epitope imprinting and nanomaterial amplification was developed. The hybrid epitope imprinting was achieved by electropolymerization in the presence of two computationally selected and cysteine modified epitopes of neuron specific enolase (NSE), as-synthesized gold nanoparticles (AuNPs), and functional monomer. The AuNPs decorated epitope-mediated hybrid MIPs, as well as the standard hybrid MIPs, were utilized for the preparation of electrochemical sensors to demonstrate the impact of nanomaterial's modification in the polymer network for biomarker sensing. The fabrication process of both sensor types was investigated by employing cyclic voltammetry, square wave voltammetry, atomic force microscopy, and scanning electron microscopy. The biomarker assay using the standard hybrid MIPs resulted in 2.5-fold higher sensitivity compared to single epitope imprints, whereas the AuNP-hybrid MIPs enhanced the sensitivity level to a great extent and allowed the recognition of NSE in human serum in a concentration range of 25-4000 pg/mL. Comparative selectivity studies with non-imprinted polymer resulted in an imprinting factor of 4.2, confirming the high target selectivity of AuNP-MIP cavities. Cross-reaction of the sensor with four reference molecules (dopamine, bovine serum albumin, glucose and elongated peptide) was negligible. As compared to current strategies for epitope imprinting which rely on single epitopes for the formation of molecular cavities, the hybrid epitope-MIPs, particularly with the inclusion of AuNPs have provided more desirable sensing platforms with high sensitivity, affinity and specificity.