New potentiometric sensor based on molecularly imprinted nanoparticles for cocaine detection

Here we present a potentiometric sensor for cocaine detection based on molecularly imprinted polymer nanoparticles (nanoMIPs) produced by the solid-phase imprinting method. The composition of polymers with high affinity for cocaine was optimised using molecular modelling. Four compositions were selected and polymers prepared using two protocols: chemical polymerisation in water and UV-initiated polymerisation in organic solvent. All synthesised nanoparticles had very good affinity to cocaine with dissociation constants between 0.6nM and 5.3nM. Imprinted polymers produced in organic solvent using acrylamide as a functional monomer demonstrated the highest yield and affinity, and so were selected for further sensor development. For this, nanoparticles were incorporated within a PVC matrix which was then used to prepare an ion-selective membrane integrated with a potentiometric transducer. It was demonstrated that the sensor was able to quantify cocaine in blood serum samples in the range of concentrations between 1nM and 1mM

Molecularly Imprinted High Affinity Nanoparticles for 4-Ethylphenol Sensing

A novel sensor for 4-ethylphenol (4EP) was developed based on molecularly imprinted nanoparticles produced via the solid phase imprinting method. The nanoparticles were immobilised on the surface of a gold electrode previously modified with a self assembled monolayer of lipoic acid coupled to EDC/NHS ((1-ethyl-3-[dimethylaminopropyl]carbodiimide)/N-hydroxysuccinimide). Each step of the process was characterised by electrochemical impedance spectroscopy and cyclic voltammetry. The electrode was used to selectively determine 4EP by differential pulse voltammetry; a quite remarkable sensitivity improvement was observed in the nanoparticle-modified electrode in reference to the non modified bare gold electrode

Formation of target-specific binding sites in enzymes: solid-phase molecular imprinting of HRP

Here we introduce a new concept for synthesising molecularly imprinted nanoparticles by using proteins as macro-functional monomers. For a proof-of-concept, a model enzyme (HRP) was cross-linked using glutaraldehyde in the presence of glass beads (solid-phase) bearing immobilized templates such as vancomycin and ampicillin. The cross-linking process links together proteins and protein chains, which in the presence of templates leads to the formation of permanent target-specific recognition sites without adverse effects on the enzymatic activity. Unlike complex protein engineering approaches commonly employed to generate affinity proteins, the method proposed can be used to produce protein-based ligands in a short time period using native protein molecules. These affinity materials are potentially useful tools especially for assays since they combine the catalytic properties of enzymes (for signaling) and molecular recognition properties of antibodies. We demonstrate this concept in an ELISA-format assay where HRP imprinted with vancomycin and ampicillin replaced traditional enzyme–antibody conjugates for selective detection of templates at micromolar concentrations. This approach can potentially provide a fast alternative to raising antibodies for targets that do not require high assay sensitivities; it can also find uses as a biochemical research tool, as a possible replacement for immunoperoxidase-conjugates