Scanning Probe and Fluorescence Analysis of Neuroreceptors Embedded in Biomimetic Membranes


Laura Blasi1, P.P. Pompa1, D. Pisignano1, G. Palazzo2, A. Mallardi3, G. Maruccio1, A. Maffei1, G. Ciccarella1, G. Vasapollo1, F. Calabi1, Roberto Cingolani1, and Ross Rinaldi1

1National Nanotechnology Laboratory of Istituto Nazionale di Fisica della Materia, Dipartimento di Ingegneria dell'Innovazione, Università di Lecce, Via per Arnesano, 73100 Lecce, Italy,
2Dipartimento di Chimica, Università di Bari, Bari, Italy, and
3Istituto per i Processi Chimico-Fisici (IPCF), CNR, Bari, Italy

e-mail: laura.blasi@unile.it

URL: http://www.nnl.it

 

The immobilization of brain dopamine receptors (primary targets in the treatment of schizophrenia, Parkinsonís disease, and Huntingtonís chorea) on a solid support can represent an appealing strategy for the realisation of biosensors for dopamine and dopaminergic drugs. Current pharmacological testing methodologies could be substituted by biosensor-based binding tests, allowing flow injection analysis, fast response and low costs. Our goal is the realization of an optical biosensor that allows a parallel quantitative analysis of different receptors, neurotransmitters and related drugs. In our system, the active layer will be constituted by dopaminergic receptors immobilized in a biomimetic membrane tethered to a suitable surface, in order to preserve biological activity.

Our goal is the realization of an optical biosensor that allows a parallel quantitative analysis of different receptors, neurotransmitters and related drugs. In our system, the active layer will be constituted by dopaminergic receptors immobilized in a biomimetic membrane tethered to a suitable surface, in order to preserve biological activity.

As tether, we chose cholesteryl derivatives, composed by a cholesterol unit attached to a polysulfide terminated diethylene glycol chain in order to provide a certain hydrophilicity in the proximity of the inorganic surface. Self-assembled monolayer (SAM) of the cholesteryl derivatives on hydrophilic surfaces were realised by means of deposition from solution and microcontact printing. The active layer was characterised by scanning probe techniques, in order to test its quality and integrity, and by fluorescence assays, using specific fluorescent ligands.