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New insights for using self-assembly materials to improve the detection stability in label-free DNA-chip and immuno-sensors
Authors:Sandro Carrara  Luca Benini  Vijayender Bhalla  Claudio Stagni  Anna Ferretti  Andrea Cavallini  Bruno Ricc  Bruno Samorì
Institution:aBiochemistry Department “G.Moruzzi”, University of Bologna, Bologna, Italy;bDepartment DEIS, University of Bologna, Bologna, Italy
Abstract:This paper examines reliable advancements in low-cost DNA- and immuno-chips. Capacitance detection was successfully chosen to develop label-free bio-chips. Probe immobilization was rigorously investigated in order to obtain reliable capacitance measurements. Protein probes immobilized by using usual alkanethiols or thiolated ssDNA probes directly immobilized on gold do not allow sufficient stable capacitance measurements. New alkanethiols improved with ethylene–glycol function are shown in this paper to be more suitable materials for capacitive bio-chip development. Atomic Force Microscopy, Quartz Crystal Microbalance, and Capacitance Measurements were used to demonstrate that ethylene–glycol alkanethiols allow high time stability, smaller errors in detection, and improved ideal behaviour of the sensing surfaces. Measured capacitance is in the range of 8–11 nF/mm2 for antibody layers and close to 6 nF/mm2 for DNA probes. It is in the range of 10–12 nF/mm2 and of 4–6 nF/mm2 for antigen and DNA detection, respectively. The percentage error in detection is highly improved and it is in the range of 11–37% and of 0,23–0,82% for antigen and DNA, respectively. The reproducibility is also improved and it is close to 0,44% for single spot measurements on ethylene–glycol alkanethiols. A molecular theory attributing these improvements to water molecules strongly coordinated by ethylene–glycol functional groups and to solution ions not entering into probe films is finally proposed.
Keywords:DNA-chip  Immuno-sensors  Capacitance detection  Alkanethiols  Ethylene–  glycol molecules  Ion layering  AFM  QCM
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