Split Aptamers Immobilized on Polymer Brushes Integrated in a Lab-on-Chip System Based on an Array of Amorphous Silicon Photosensors: A Novel Sensor Assay

Manasa Nandimandalam; Francesca Costantini; Nicola Lovecchio; Lorenzo Iannascoli; Augusto Nascetti; Giampiero de Cesare; Domenico Caputo; Cesare Manetti

doi:10.3390/ma14237210

Split Aptamers Immobilized on Polymer Brushes Integrated in a Lab-on-Chip System Based on an Array of Amorphous Silicon Photosensors: A Novel Sensor Assay

Materials (Basel). 2021 Nov 26;14(23):7210. doi: 10.3390/ma14237210.

Authors

Manasa Nandimandalam¹, Francesca Costantini^{1

2}, Nicola Lovecchio³, Lorenzo Iannascoli^{3

4}, Augusto Nascetti⁴, Giampiero de Cesare³, Domenico Caputo³, Cesare Manetti¹

Affiliations

¹ Department of Environmental Biology, Sapienza University of Rome, P.le Aldo Moro 5, 00185 Rome, Italy.
² CREA-DC Research Centre for Plant Protection and Certification, 00156 Rome, Italy.
³ Department of Information, Electronic and Telecommunication Engineering, Sapienza University of Rome, Via Eudossiana 18, 00184 Roma, Italy.
⁴ School of Aerospace Engineering, Sapienza University of Rome, Via Salaria 851/881, 00138 Rome, Italy.

Abstract

Innovative materials for the integration of aptamers in Lab-on-Chip systems are important for the development of miniaturized portable devices in the field of health-care and diagnostics. Herein we highlight a general method to tailor an aptamer sequence in two subunits that are randomly immobilized into a layer of polymer brushes grown on the internal surface of microfluidic channels, optically aligned with an array of amorphous silicon photosensors for the detection of fluorescence. Our approach relies on the use of split aptamer sequences maintaining their binding affinity to the target molecule. After binding the target molecule, the fragments, separately immobilized to the brush layer, form an assembled structure that in presence of a "light switching" complex [Ru(phen)₂(dppz)]²⁺, emit a fluorescent signal detected by the photosensors positioned underneath. The fluorescent intensity is proportional to the concentration of the target molecule. As proof of principle, we selected fragments derived from an aptamer sequence with binding affinity towards ATP. Using this assay, a limit of detection down to 0.9 µM ATP has been achieved. The sensitivity is compared with an assay where the original aptamer sequence is used. The possibility to re-use both the aptamer assays for several times is demonstrated.

Keywords: ATP; Lab-on-Chip; a-Si:H photosensors; polymer brushes; split aptamers.