Liebich, Valentina Jutta (2023)
Engineering of Bioactive Papers: Site-specific, Covalent Immobilization of Peptides and Proteins.
Technische Universität Darmstadt
doi: 10.26083/tuprints-00022853
Dissertation, Erstveröffentlichung, Verlagsversion
Kurzbeschreibung (Abstract)
Bioactive papers are of growing interest regarding point-of-care testing devices that do not require extensive analytical equipment and have with Covid-19 made their way to everyone’s household. The generation of such papers that comprise - in their simplest form - a bioactive protein presented on a cellulosic material, typically relies on four very different immobilization principles, which can be divided into covalent/non-covalent and site-specific/non-site-specific strategies. Covalent, site-specific approaches are of special demand if a uniform, site-directed distribution of the biomolecule throughout the paper substrate is required along with a stable binding. Modular, chemoenzymatic approaches enable the formation of such bonds. To the best of our knowledge, we are the first to investigate chemoenzymatic approaches for functional protein immobilization on cotton linters-based materials aimed at a versatile toolbox of strategies for the efficient and reliable generation of bioactive papers. The synthetic peptide immobilization strategies rely on: 1) highly efficient, spatially controllable phototriggered cycloaddition, 2) near to “green” oxime ligation, 3) fast reductive amination with subsequent amidation, or 4) straightforward esterification with subsequent amidation. All approaches are followed by site-specific sortase A- or microbial transglutaminase-catalyzed transamidation. These four different conjugation strategies for peptide immobilization were evaluated with respect to reproducibility and fiber loading efficiency. The two enzymes employed for protein conjugation were able to recognize the same oligoglycine peptide anchor. Both proved suitable for controlled and site-directed conjugation of substrate proteins at physiological conditions. The method was engineered to allow unidirectional and covalent immobilization of several proteins displaying different functional properties, with ramifications for application in paper-based diagnostics.
Typ des Eintrags: | Dissertation | ||||
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Erschienen: | 2023 | ||||
Autor(en): | Liebich, Valentina Jutta | ||||
Art des Eintrags: | Erstveröffentlichung | ||||
Titel: | Engineering of Bioactive Papers: Site-specific, Covalent Immobilization of Peptides and Proteins | ||||
Sprache: | Englisch | ||||
Referenten: | Kolmar, Prof. Dr. Harald ; Biesalski, Prof. Dr. Markus | ||||
Publikationsjahr: | 2023 | ||||
Ort: | Darmstadt | ||||
Kollation: | x, 128 Seiten | ||||
Datum der mündlichen Prüfung: | 31 Oktober 2022 | ||||
DOI: | 10.26083/tuprints-00022853 | ||||
URL / URN: | https://tuprints.ulb.tu-darmstadt.de/22853 | ||||
Kurzbeschreibung (Abstract): | Bioactive papers are of growing interest regarding point-of-care testing devices that do not require extensive analytical equipment and have with Covid-19 made their way to everyone’s household. The generation of such papers that comprise - in their simplest form - a bioactive protein presented on a cellulosic material, typically relies on four very different immobilization principles, which can be divided into covalent/non-covalent and site-specific/non-site-specific strategies. Covalent, site-specific approaches are of special demand if a uniform, site-directed distribution of the biomolecule throughout the paper substrate is required along with a stable binding. Modular, chemoenzymatic approaches enable the formation of such bonds. To the best of our knowledge, we are the first to investigate chemoenzymatic approaches for functional protein immobilization on cotton linters-based materials aimed at a versatile toolbox of strategies for the efficient and reliable generation of bioactive papers. The synthetic peptide immobilization strategies rely on: 1) highly efficient, spatially controllable phototriggered cycloaddition, 2) near to “green” oxime ligation, 3) fast reductive amination with subsequent amidation, or 4) straightforward esterification with subsequent amidation. All approaches are followed by site-specific sortase A- or microbial transglutaminase-catalyzed transamidation. These four different conjugation strategies for peptide immobilization were evaluated with respect to reproducibility and fiber loading efficiency. The two enzymes employed for protein conjugation were able to recognize the same oligoglycine peptide anchor. Both proved suitable for controlled and site-directed conjugation of substrate proteins at physiological conditions. The method was engineered to allow unidirectional and covalent immobilization of several proteins displaying different functional properties, with ramifications for application in paper-based diagnostics. |
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Alternatives oder übersetztes Abstract: |
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Status: | Verlagsversion | ||||
URN: | urn:nbn:de:tuda-tuprints-228539 | ||||
Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 500 Naturwissenschaften und Mathematik > 500 Naturwissenschaften 500 Naturwissenschaften und Mathematik > 540 Chemie 500 Naturwissenschaften und Mathematik > 570 Biowissenschaften, Biologie |
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Fachbereich(e)/-gebiet(e): | 07 Fachbereich Chemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie 07 Fachbereich Chemie > Clemens-Schöpf-Institut > Fachgebiet Biochemie > Allgemeine Biochemie |
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Hinterlegungsdatum: | 14 Feb 2023 13:05 | ||||
Letzte Änderung: | 15 Feb 2023 07:54 | ||||
PPN: | |||||
Referenten: | Kolmar, Prof. Dr. Harald ; Biesalski, Prof. Dr. Markus | ||||
Datum der mündlichen Prüfung / Verteidigung / mdl. Prüfung: | 31 Oktober 2022 | ||||
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