Optical sensing of phenylalanine in urine via extraction with magnetic molecularly imprinted poly(ethylene-co-vinyl alcohol) nanoparticles.

Incorporation of superparamagnetic nanoparticles into molecularly imprinted polymers (MIPs) is useful for both bioseparations and for concentration and sensing of biomedically relevant target molecules in physiological fluids, through the application of a magnetic field. In this study, we combined the separation and concentration of a target (phenylalanine) in urine, using magnetic molecularly imprinted polymeric composite nanoparticles, with optical sensing, to improve assay sensitivity. This target is important as a catecholamine precursor, and as an important amino acid constituent of proteins. Poly(ethylene-co-vinyl alcohol)s were imprinted with target molecules, and showed a high imprinting effectiveness (target binding compared with binding to non-imprinted polymer particles.) Fluorescence spectrophotometry was used to measure binding of the target, and also binding of possible interfering compounds. These measurements suggest that functional groups on phenylalanine dominate the selectivity of the synthesized MIPs. Finally, the composite nanoparticles were used to separate and sense the target molecule in urine by Raman scattering microscopy.

Microcontact imprinting of algae for biofuel systems: the effects of the polymer concentration.

Microcontact imprinting of cells often involves the deposition of a polymer solution onto a monolayer cell stamp, followed by solvent evaporation. Thus, the concentration of the polymer may play an important role in the final morphology and efficacy of the imprinted film. In this work, various concentrations of poly(ethylene-co-vinyl alcohol) (EVAL) were dissolved in dimethyl sulfoxide (DMSO) for the microcontact imprinting of algae on an electrode. Scanning electron microscopy and fluorescence spectrometry were used to characterize the surface morphology and recognition capacity of algae to the algae-imprinted cavities. The readsorption of algae onto algae-imprinted EVAL thin films was quantified to obtain the EVAL concentration that maximized algal binding. Finally, the power and current density of an algal biofuel cell with the algae-imprinted EVAL-coated electrode were measured and found to be approximately double those of such a cell with a Pt/indium tin oxide (ITO)/poly(ethylene terephthalate) (PET) electrode.

Recognition of Rhodobacter sphaeroides by microcontact-imprinted poly(ethylene-co-vinyl alcohol).

The immobilization of cells or microorganisms is important for bioseparations, in bioreactors producing cellular metabolites, and as receptors for biosensing. Cell-imprinted polymers (CIPs) have been shown to have cavities with complementary shapes and also high affinities for the template cells or microorganisms. However, the effects of binding to CIPs on gene expression are only beginning to be studied. In this work, the purple bacteria Rhodobacter sphaeroides was employed as a model for the imprinting of microorganisms. R. sphaeroides was first adsorbed on a glass slide as the stamp and then microcontact-imprinted onto poly(ethylene-co-vinyl alcohol), EVAL. The surfaces of the R. sphaeroides-imprinted (RsIPs) and non-imprinted (NIPs) EVAL thin films were examined by Raman spectrometry and scanning electron microscopy. The expression of the nitrogenase (nitrogen fixation, nifH) gene of R. sphaeroides adsorbed on both the RsIPs and NIPs EVAL thin films was also measured by the quantitative reverse transcription polymerase chain reaction (qRT-PCR); cells grown on imprinted polymer showed dramatic differences in gene expression compared to controls.