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UV resonance Raman spectroscopy of artificial supramolecular ligands with laser excitation wavelengths between 244 - 300 nm
Ultraviolet resonance Raman (UVRR) has been proven to be a powerful vibrational spectroscopic tool for a selective, sensitive, and label-free monitoring of peptides and proteins by artificial supramolecular ligands like guanidiniocarbonyl pyrroles (GCPs). However, so far the use of UVRR spectroscopy is limited by the UV-excited autofluorescence of the aromatic amino acids or even from the ligand itself which can mask the UVRR signal. Two ways to remedy the autofluorescence issue are explored with the help of a pulsed tunable laser system; spectral separation by shifting the excitation wavelength and temporal separation by a so called Kerr gate. Three different artificial ligands were studied, the two oxoanion binders GCP-ethylamide and its analogue guanidinocarbonyl indole (GCI) ethylamide and the molecular tweezers CLR01 which binds selectively to lysine and arginine. The molecular tweezers signify an attempt to extend the scope of UVRR spectroscopy to a new class of supramolecular ligands. An UVRR spectrum of CLR01 could be acquired, however, the Raman signal of the phosphate groups, which are significant for binding, are not enhanced. For GCP-ethylamide an excitation study was performed between 244~nm and 310~nm to determine the laser excitation wavelength for an optimal UVRR signal with minimal impeding autofluorescence. The GCP analogue GCI exhibits a resonance around 244~nm which allows for autofluorescence-free acquisition of UVRR spectra. Spectral changes upon binding are first shown for two test molecules: the aromatic benzoic acid and the ubiquitous tripeptide RGD (arginylglycylaspartic acid). First tests were also carried out with the proteins bovine serum albumin (BSA) and Survivin. For a more generic approach a Kerr gate operating in the UV spectral range was set up. The separation of UVRR signal from autofluorescence is independent from the used laser excitation wavelength and can be applied for all molecules. The Kerr gate is demonstrated to work for a laser excitation wavelength of 280~nm. All ...
UV resonance Raman spectroscopy of artificial supramolecular ligands with laser excitation wavelengths between 244 - 300 nm
Ultraviolet resonance Raman (UVRR) has been proven to be a powerful vibrational spectroscopic tool for a selective, sensitive, and label-free monitoring of peptides and proteins by artificial supramolecular ligands like guanidiniocarbonyl pyrroles (GCPs). However, so far the use of UVRR spectroscopy is limited by the UV-excited autofluorescence of the aromatic amino acids or even from the ligand itself which can mask the UVRR signal. Two ways to remedy the autofluorescence issue are explored with the help of a pulsed tunable laser system; spectral separation by shifting the excitation wavelength and temporal separation by a so called Kerr gate. Three different artificial ligands were studied, the two oxoanion binders GCP-ethylamide and its analogue guanidinocarbonyl indole (GCI) ethylamide and the molecular tweezers CLR01 which binds selectively to lysine and arginine. The molecular tweezers signify an attempt to extend the scope of UVRR spectroscopy to a new class of supramolecular ligands. An UVRR spectrum of CLR01 could be acquired, however, the Raman signal of the phosphate groups, which are significant for binding, are not enhanced. For GCP-ethylamide an excitation study was performed between 244~nm and 310~nm to determine the laser excitation wavelength for an optimal UVRR signal with minimal impeding autofluorescence. The GCP analogue GCI exhibits a resonance around 244~nm which allows for autofluorescence-free acquisition of UVRR spectra. Spectral changes upon binding are first shown for two test molecules: the aromatic benzoic acid and the ubiquitous tripeptide RGD (arginylglycylaspartic acid). First tests were also carried out with the proteins bovine serum albumin (BSA) and Survivin. For a more generic approach a Kerr gate operating in the UV spectral range was set up. The separation of UVRR signal from autofluorescence is independent from the used laser excitation wavelength and can be applied for all molecules. The Kerr gate is demonstrated to work for a laser excitation wavelength of 280~nm. All ...
UV resonance Raman spectroscopy of artificial supramolecular ligands with laser excitation wavelengths between 244 - 300 nm
Holtum, Tim Henrik (author) / Schlücker, Sebastian
2022-06-07
Theses
Electronic Resource
English
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