New micro-and nano-technologies for biosensor development

Recent advances in micro- and nanotechnology have produced a number of new materials which exhibit exceptional potential for the design of novel sensing strategies and to enhance the analytical performance of biosensing systems. In this thesis three different types of miniaturisation pathways were investigated for electrochemical biosensing applications. Vertically aligned carbon nanotube thin films were designed and tested as platforms for DNA immobilisation and for the development of a model electrochemical genosensor. The sensor format involved the immobilisation of oligoucleotide probes onto the sensor surface, hybridisation with the target sequence and electrochemical detection of the duplex formation. By combining such an electrode platform with an enzyme labeling, a detection limit of oligonucleotide targets in the nanomolar range was achieved. A novel magnetic particle-based microfluidic sensor was also realised by integrating a microfluidic platform with a new analytical procedure based on the use of paramagnetic beads for the detection of real PCR samples. The hybridisation reaction was carried out on probe-modified beads in a flow-through format, thus enhancing the surface area-to-volume ratio and consequently the sensitivity. Moreover, the magnetic properties of the beads greatly facilitated the delivery and removal of reagents through the microfluidic channels. This format allowed the detection of nanomolar levels of double-stranded DNA sequences, with high reproducibility and fast time of analysis. Finally, polyaniline nanotubes arranged in an ordered structure directly on gold electrode surfaces were realised and employed to create a model molecularly imprinted (MIP) polymer -sensor for catechol detection. The advantages of using nanostructures in this particular biosensing application have been evaluated by comparing the analytical performance of the sensor with an analogous non-nanostructured MIP-sensor that we had previously developed. A significantly lower limit of detection (one order of magnitude) was achieved, thus

demonstrating that the nanostructures enhanced the analytical performance of the sensor.