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Identifier 000451006
Title Development of a combined surface plasmon resonance and Love wave - surface acoustic wave system for biophysical studies
Alternative Title Ανάπτυξη συστήματος που συνδυάζει τις τεχνικές συντονισμού επιφανειακών πλασμονίων και επιφανειακών ακουστικών κυμάτων τύπου Love για βιοφυσικές μελέτες
Author Σαμαρεντσής, Αναστάσιος Γ
Thesis advisor Γκιζελή, Ηλέκτρα
Reviewer Francis, Laurent A.
Friedt, Jean Michel
Χαλεπάκης, Γιώργος
Αθανασάκη, Ειρήνη
Gogolides, Evangelos
Mitsakakis, Konstantinos
Abstract The field of biosensing is closely related to the development of sensitive analytical techniques. Biosensors can be defined as such tools, incorporating a sensitive biological element and a transducer for the detection of a target analyte. Despite the great advances in the field of biosensors, there is still an ongoing demand for more accurate analytical information. In this PhD research project, we present the development of a novel combined surface plasmon resonance (SPR) and Love wave (LW) surface acoustic wave (SAW) system for biophysical studies. To achieve the above, we first investigated the experimental parameters required to match simultaneously the conditions for efficient SPR coupling and acoustic waveguiding. A hybrid SPR/LW-SAW device was designed and fabricated incorporating a PMMA waveguide layer and a Au-coated sensing area. The sensor was evaluated in terms of materials and structures in order to identify the optimal thickness for the PMMA and Au layer. In addition to that, a fluidic system consisting of a 3D-printed plastic holder integrated with a PDMS microfluidic chip was developed to allow experiments under continuous flow. The system was calibrated in a systematic study of the optical and acoustic responses for different surface perturbations i.e., rigid mass loading (Au deposition) and pure viscous loading (glycerol and sucrose solutions). Subsequently, simultaneous optical and acoustic measurements were employed for monitoring biomolecular interactions on a single surface. First, the combined SPR/LW-SAW platform was used to study the adsorption of proteins on Au. The proteins tested include bovine serum albumin (BSA) and three biotin-binding proteins; avidin (Av), streptavidin (SAv) and neutravidin (NAv). We proved that through appropriate quantitative interpretation of the combined data, it is feasible to extract protein film properties not accessible with a single technique. Finally, the system was applied for the characterization of double-stranded DNA molecules (dsDNAs) of various sizes immobilized on a SAv-modified, Au surface through a SAv-biotin link. Acoustic and optical signals were analyzed to get further insight into the properties of the immobilized DNA molecules including surface mass, conformation status and water content. Having verified the abovementioned characteristics, we believe that the developed platform could lead to improved understanding of dynamic processes at biointerfaces.
Language English
Subject Biosensors
Biotin-binding proteins (Avidin, Streptavidin and Neutravidin)
Combined SPR/LW-SAW biosensing platform
Double-stranded DNA molecules (DSDNA)
Protein adsorption
Βιοαισθητήρες
Βιοφυσική μελέτη
Επιφανειακά ακουστικά κύματα τύπου Love (LW-SAW)
Προσρόφηση πρωτεϊνών
Πρωτεΐνες που προσδένουν Βιοτίνη (Αβιδίνη, Στρεπταβιδίνη και Νιουτραβιδίνη)
Συντονισμός επιφανειακών πλασμονίων (SPR)
Χαρακτηρισμός μορίων δίκλωνου DNA (DSDNA)
Issue date 2022-10-07
Collection   School/Department--School of Sciences and Engineering--Department of Biology--Doctoral theses
  Type of Work--Doctoral theses
Permanent Link https://elocus.lib.uoc.gr//dlib/1/e/b/metadata-dlib-1662720880-104933-3708.tkl Bookmark and Share
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