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- ItemErarbeitung eines mobilen mikrosystemtechnischen Gesamtsystems für Anreicherung, Nachweis und Charakterisierung zirkulierender Tumorzellen, Teilvorhaben IPHT: Entwicklung eines chipbasierten elektrischen Nachweises zur parallelen Identifikation von Tumorspezifischen Markern : Abschlussbericht ; Laufzeit des Vorhabens: 01.01.2011 - 30.06.2014(Hannover : Technische Informationsbibliothek (TIB), 2014) Weber, Karina; Popp, JürgenDie Charakterisierung von zirkulierenden Tumorzellen stellt ein wichtiges diagnostisches Hilfsmittel für das Therapiemonitoring und die Therapieoptimierung bei Krebspatienten dar. Im Fokus des Verbundvorhabens „MiNa-CTC“ stand daher die Erforschung und Realisierung eines miniaturisierten, kostengünstigen, zuverlässigen, effizienten und mobilen Gesamtsystems für die Anreicherung, den Nachweis und die Charakterisierung zirkulierender Tumorzellen. Das Teilvorhaben des IPHT befasst sich dabei insbesondere mit der Erforschung einer chipbasierten Nachweismethode zur Detektion verschiedener Tumor-spezifischer Marker. Hierbei wurden insbesondere die Biomarker KRAS, PIK3CA und BRAF sowie im Hinblick auf eine mögliche Real-Time Amplifikation die Referenzgene RPL13A und GusBeta betrachtet. Im Ergebnis steht eine optisch auslesbare und miniaturisierte Chip-Plattform zum spezifischen Nachweis von Punktmutationen, die als Teillösung in ein Gesamtsystem integriert werden konnte. Das modular aufgebaute Gesamtsystem konnte erfolgreich an verschiedenen Probenmatrices validiert werden und stellt die Basis für ein zukünftig dezentral einsetzbares System zum Therapiemonitoring dar.
- ItemIn Vitro Selection of Specific DNA Aptamers Against the Anti-Coagulant Dabigatran Etexilate(Berlin : Nature Publishing, 2018) Aljohani, Maher M; Chinnappan, Raja; Eissa, Shimaa; Alsager, Omar A; Weber, Karina; Cialla-May, Dana; Popp, Jürgen; Zourob, MohammedDabigatran Etexilate (PRADAXA) is a new oral anticoagulant increasingly used for a number of blood thrombosis conditions, prevention of strokes and systemic emboli among patients with atrial fibrillation. It provides safe and adequate anticoagulation for prevention and treatment of thrombus in several clinical settings. However, anticoagulation therapy can be associated with an increased risk of bleeding. There is a lack of specific laboratory tests to determine the level of this drug in blood. This is considered the most important obstacles of using this medication, particularly for patients with trauma, drug toxicity, in urgent need for surgical interventions or uncontrolled bleeding. In this work, we performed Systematic evolution of ligands by exponential enrichment (SELEX) to select specific DNA aptamers against dabigatran etexilate. Following multiple rounds of selection and enrichment with a randomized 60-mer DNA library, specific DNA aptamers for dabigatran were selected. We investigated the affinity and specificity of generated aptamers to the drug showing dissociation constants (Kd) ranging from 46.8–208 nM. The most sensitive aptamer sequence was selected and applied in an electrochemical biosensor to successfully achieve 0. 01 ng/ml level of detection of the target drug. With further improvement of the assay and optimization, these aptamers would replace conventional antibodies for developing detection assays in the near future.Dabigatran Etexilate (PRADAXA) is a new oral anticoagulant increasingly used for a number of blood thrombosis conditions, prevention of strokes and systemic emboli among patients with atrial fibrillation. It provides safe and adequate anticoagulation for prevention and treatment of thrombus in several clinical settings. However, anticoagulation therapy can be associated with an increased risk of bleeding. There is a lack of specific laboratory tests to determine the level of this drug in blood. This is considered the most important obstacles of using this medication, particularly for patients with trauma, drug toxicity, in urgent need for surgical interventions or uncontrolled bleeding. In this work, we performed Systematic evolution of ligands by exponential enrichment (SELEX) to select specific DNA aptamers against dabigatran etexilate. Following multiple rounds of selection and enrichment with a randomized 60-mer DNA library, specific DNA aptamers for dabigatran were selected. We investigated the affinity and specificity of generated aptamers to the drug showing dissociation constants (Kd) ranging from 46.8–208 nM. The most sensitive aptamer sequence was selected and applied in an electrochemical biosensor to successfully achieve 0. 01 ng/ml level of detection of the target drug. With further improvement of the assay and optimization, these aptamers would replace conventional antibodies for developing detection assays in the near future.