Randy De Palma

Stability of mixed PEO-thiol SAMs for biosensing applications.

The secret of a successful affinity biosensor partially hides in the chemical interface layer between the transducer system and the biological receptor molecules. Over the past decade, several methodologies for the construction of such interface layers have been developed on the basis of the deposition of self-assembled monolayers (SAMs) of alkanethiols on gold. Moreover, mixed SAMs of polyethylene oxide (PEO) containing thiols have been applied for the immobilization of biological receptors. Despite the intense research in the field of thiol SAMs, relatively little is known about their biosensing properties in correlation with their long-term stability. Especially the impact of the storage conditions on their biosensing characteristics has not been reported before to our knowledge. To address these issues, we prepared mixed PEO SAMs and tested their stability and biosensing performance in several storage conditions, i.e., air, N2, ethanol, phosphate buffer, and H2O. The quality of the SAMs was monitored as a function of time using various characterization techniques such as cyclic voltammetry, contact angle, grazing angle Fourier-transform infrared spectroscopy, and X-ray photoelectron spectroscopy. In addition, the impact of the different storage conditions on the biosensor properties was investigated using surface plasmon resonance. Via the latter technique, the receptor immobilization, the analyte recognition, and the nonspecific binding were extensively studied using the prostate specific antigen as a model system. Our experiments showed that very small structural differences in the SAM can have a great impact in their final biosensing properties. In addition it was shown that the mixed SAMs stored in air or N2 are very stable and retain their biosensor properties for at least 30 days, while ethanol appeared to be the worst storage medium due to partial oxidation of the thiol headgroup. In conclusion, care must be taken to avoid SAM degradation during storage to retain typical SAM characteristics, which is very important for their general use in many proposed applications.

Discrimination of specific and non-specific bindings by dielectrophoretic repulsion in on-chip magnetic bio-assays.

Affinity binding is the principle used in a large number of bio-assays. Aside from specific bindings, non-specific bindings usually deteriorate assays by giving false positive signals and restrict the detection limit. Currently, the assay specificity is mainly dependent on the effectiveness of a suitable surface chemistry. We report an approach to discriminate specific and non-specific bindings with dielectrophoretic (DEP) forces for on-chip magnetic bio-assays. Conjugated to the analytes, magnetic particles were used as the agents for DEP force generation. Due to a weaker binding strength, the non-specifically bound particles were removed while specific bindings remained intact. Analytical and finite element calculations were also performed to study all relevant forces. Furthermore, the removal of magnetic particles was also assessed by measuring the magnetic signal using magnetoresistive sensors. This technique can not only be used to improve the specificity of the on-chip bio-assays but also be developed as a tool of force spectroscopy for the study of bio-molecular binding physics.