RESUMO
Bioactive protein patterns and microarrays achieved by selective localization of biomolecules find various applications in biosensors, bio-microelectromechanical systems (bio-MEMS), and in basic protein studies. In this paper we describe simple photochemical methods to fabricate two-dimensional patterns on a Novolac A derivative polymer (SU-8) and, subsequently, their functionalization with biomolecules. Anthraquinone (AQ) derivatives are used to chemically modify and pattern SU-8 surfaces. Features as small as 20 mum are obtained when using uncollimated light. The X-Y spatial resolution of micropatterned AQ molecules is improved to 1.5 mum when a collimated light source is used. This micropatterning process will be important for the functionalization of MEMS-based biosensors. The method saves several processing steps and can be integrated in cleanroom fabrication thus avoiding contamination of the sensor surfaces.
Assuntos
Antraquinonas/farmacologia , Corantes/farmacologia , Compostos de Epóxi/farmacologia , Fotoquímica/métodos , Polímeros/farmacologia , Análise Serial de Proteínas/métodos , Adsorção , Técnicas Biossensoriais , Reagentes de Ligações Cruzadas/farmacologia , Compostos de Epóxi/química , Luz , Modelos Químicos , Polímeros/química , Proteínas/química , Espectrometria de Fluorescência/métodos , Propriedades de Superfície , Fatores de TempoRESUMO
Here, we present the activities within our research group over the last five yearswith cantilevers fabricated in the polymer SU-8. We believe that SU-8 is an interestingpolymer for fabrication of cantilevers for bio/chemical sensing due to its simple processingand low Young's modulus. We show examples of different integrated read-out methodsand their characterisation. We also show that SU-8 cantilevers have a reduced sensitivity tochanges in the environmental temperature and pH of the buffer solution. Moreover, weshow that the SU-8 cantilever surface can be functionalised directly with receptormolecules for analyte detection, thereby avoiding gold-thiol chemistry.
RESUMO
This paper describes the development of novel particle-based fluorescence resonance energy transfer (FRET) sensors to quantify the concentration and monitor the binding affinity of carbohydrates and glycoproteins to lectins, which are carbohydrate binding proteins. The sensing approach is based on FRET between fluorescein (donor)-labeled lectin molecules, adsorbed on the surface of micrometric polymeric beads, and polymeric dextran molecules labeled with Texas Red (acceptor). The FRET efficiency of the donor-acceptor pair decreases in the presence of carbohydrates or glycoproteins that compete with the Texas Red-labeled dextran molecules on the lectinic binding sites. The inhibitory effect is concentration and time dependent. The sensing technique enables the discrimination between carbohydrates and glycoproteins based on their binding affinity to the FRET sensing particles as well as quantitative analysis of carbohydrates and glycoproteins in aqueous samples. In the future, the newly developed sensors could enable screening glycoprotein-based drugs for their binding affinity toward selective receptors.