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Development of bioanalytical assays on LSPR-based sensors
This work is focused on LSPR-based bioanalysis using low-cost sensor devices with chemically synthesized gold nanoparticles as transducer for bioassays in single and array formats. Interactions of analyte molecules are investigated both directly on the surface of gold nanoparticles and on differently immobilized capture molecules. Various solutions like glycerol of different concentrations and layer by layer depositions of positively and negatively charged polyelectrolytes are used to determine the bulk and surface sensitivities of the sensors. To determine the optimal conditions for biomolecule detection, the specific interactions of proteins and DNA molecules are investigated. For protein detection, the effects of different immobilization methods of capture molecules leading to different distances of analyte binding motifs to the gold surface on the analyte signal were demonstrated. It has been shown that toxicity-relevant studies of protein interactions at the surface of gold nanoparticles can be observed similarly well with the LSPR instrument as, for example, with the well-established method of fluorescence spectroscopy. However, the signal strength for specific protein detections decreases with the distance of the analyte binding motif (capture molecule) to the gold surface as expected. Nevertheless, the presented system is able to achieve detection limits for CRP in the clinically relevant range even at the largest distance. The effect of analytes of different sizes on their LSPR signal was investigated using DNA molecules. The result was that the shortest analytes performed best (highest signal), but this is probably due to steric hindrance by the larger molecules. In addition, analytes with single base mismatches could also be distinguished from the fully complementary sequence using the established low-cost system. Finally, the optimization of plasmonic array sensors using a newly developed readout device was demonstrated. The homogeneity of the arrays could be significantly improved by adjusting the ...
Development of bioanalytical assays on LSPR-based sensors
This work is focused on LSPR-based bioanalysis using low-cost sensor devices with chemically synthesized gold nanoparticles as transducer for bioassays in single and array formats. Interactions of analyte molecules are investigated both directly on the surface of gold nanoparticles and on differently immobilized capture molecules. Various solutions like glycerol of different concentrations and layer by layer depositions of positively and negatively charged polyelectrolytes are used to determine the bulk and surface sensitivities of the sensors. To determine the optimal conditions for biomolecule detection, the specific interactions of proteins and DNA molecules are investigated. For protein detection, the effects of different immobilization methods of capture molecules leading to different distances of analyte binding motifs to the gold surface on the analyte signal were demonstrated. It has been shown that toxicity-relevant studies of protein interactions at the surface of gold nanoparticles can be observed similarly well with the LSPR instrument as, for example, with the well-established method of fluorescence spectroscopy. However, the signal strength for specific protein detections decreases with the distance of the analyte binding motif (capture molecule) to the gold surface as expected. Nevertheless, the presented system is able to achieve detection limits for CRP in the clinically relevant range even at the largest distance. The effect of analytes of different sizes on their LSPR signal was investigated using DNA molecules. The result was that the shortest analytes performed best (highest signal), but this is probably due to steric hindrance by the larger molecules. In addition, analytes with single base mismatches could also be distinguished from the fully complementary sequence using the established low-cost system. Finally, the optimization of plasmonic array sensors using a newly developed readout device was demonstrated. The homogeneity of the arrays could be significantly improved by adjusting the ...
Development of bioanalytical assays on LSPR-based sensors
Kastner, Stephan (author) / Neugebauer, Ute / Fritzsche, Wolfgang
2023-01-01
Theses
Electronic Resource
English
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