Cleavable probes and catch and release assays for surface group quantification on 2D- and 3D-supports
- Surface functionalization of 2D- and 3D-supports and nanomaterials are nowadays at the core of many applications of functional materials in the life and material sciences. Examples range from membranes and microarrays over bead-based assays, immunoseparation, and next generation sequencing to nanometer-sized optical reporters, nanosensors, and magnetic and optical contrast agents. Typically performed functionalization procedures include silanization and grafting reactions with reactive monomers to introduce functional groups like amino or carboxylic acid groups and the attachment of ligands like polyethylene glycol (PEG) molecules and biomolecules such as peptides, proteins, and DNA.[1-3] We present here a versatile concept to quantify the number of bioanalytically relevant functional groups like carboxyl, amino, and aldehyde moieties through the specific binding and subsequent release of small reporter molecules such as fluorescent dyes and non-fluorescent chromophores utilizingSurface functionalization of 2D- and 3D-supports and nanomaterials are nowadays at the core of many applications of functional materials in the life and material sciences. Examples range from membranes and microarrays over bead-based assays, immunoseparation, and next generation sequencing to nanometer-sized optical reporters, nanosensors, and magnetic and optical contrast agents. Typically performed functionalization procedures include silanization and grafting reactions with reactive monomers to introduce functional groups like amino or carboxylic acid groups and the attachment of ligands like polyethylene glycol (PEG) molecules and biomolecules such as peptides, proteins, and DNA.[1-3] We present here a versatile concept to quantify the number of bioanalytically relevant functional groups like carboxyl, amino, and aldehyde moieties through the specific binding and subsequent release of small reporter molecules such as fluorescent dyes and non-fluorescent chromophores utilizing cleavable linkers or the formation of cleavable bonds as a reversible covalent labeling strategy. This is representatively demonstrated for different types of nano- and microparticles with different labeling densities of carboxyl, amino, and aldehyde groups. This strategy enables to separate the signal-generating molecule from the bead surface, thereby circumventing uncertainties associated with light scattering, binding-induced changes in reporter fluorescence, and fluorescence quenching dye-dye interactions on crowded material surfaces.[1-3] Moreover, the reporters are chosen to be detectable with different analytical methods as prerequisite for straightforward validation via method compari-sons and mass balances. Applications of these assays and multimodal cleavable probes range from a quantitative comparison of bead batches and process control to a qualitative prediction of the coupling efficiencies in bioconjugation reactions.…
Autor*innen: | Ute Resch-GengerORCiD |
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Koautor*innen: | Nithiya Nirmalananthan-Budau, Marko Moser, Alexander Roloff, Bastian RühleORCiD, H. Borcherding, T. Thiele, U. Schedler |
Dokumenttyp: | Vortrag |
Veröffentlichungsform: | Präsentation |
Sprache: | Englisch |
Jahr der Erstveröffentlichung: | 2019 |
Organisationseinheit der BAM: | 1 Analytische Chemie; Referenzmaterialien |
1 Analytische Chemie; Referenzmaterialien / 1.2 Biophotonik | |
DDC-Klassifikation: | Technik, Medizin, angewandte Wissenschaften / Ingenieurwissenschaften / Ingenieurwissenschaften und zugeordnete Tätigkeiten |
Freie Schlagwörter: | Cleavable probe; Conductometry; Dye labeling; Methos comparision; Nanoparticle; Optical assay; Quality assurance; Surface analysis; Surface group |
Themenfelder/Aktivitätsfelder der BAM: | Material |
Material / Nano | |
Veranstaltung: | ANAKON 2019 |
Veranstaltungsort: | Münster, Germany |
Beginndatum der Veranstaltung: | 25.03.2019 |
Enddatum der Veranstaltung: | 28.03.2019 |
Verfügbarkeit des Dokuments: | Datei im Netzwerk der BAM verfügbar ("Closed Access") |
Datum der Freischaltung: | 27.03.2019 |
Referierte Publikation: | Nein |
Eingeladener Vortrag: | Nein |