Publikationsserver der Universitätsbibliothek Marburg

Titel:Neue Ansätze zur Entwicklung eines Ganzzellbiosensors
Autor:Vornholt, Wolfgang
Weitere Beteiligte: Keusgen, Michael (Prof. Dr.)
Veröffentlicht:2010
URI:https://archiv.ub.uni-marburg.de/diss/z2010/0458
DOI: https://doi.org/10.17192/z2010.0458
URN: urn:nbn:de:hebis:04-z2010-04586
DDC: Naturwissenschaften
Titel(trans.):New approaches for the development of a whole cell biosensor
Publikationsdatum:2010-07-08
Lizenz:https://rightsstatements.org/vocab/InC-NC/1.0/

Dokument

Schlagwörter:
Oberflächenplasmonresonanz, Lektine, Lectins, Lipopolysaccharid, Lipopolysaccharide

Zusammenfassung:
Unter Verwendung der Oberflächenplasmonresonanz (SPR) wurden unterschiedliche Modellapplikationen etabliert. Es wurden verschiedene Zuckeroberflächen erzeugt und durch ein Lektin-Screening charakterisiert. Das Bindungsverhalten derartiger Oberflächen wurde daraufhin an NCI-H125- sowie an Lewis Lung-Zellen untersucht. Im Hinblick auf eine Mistellektinbestimmung in Fertigarzneimitteln zur Krebstherapie wurde ein Mistellektin-Assay entwickelt. Die Detektion des rekombinanten Mistellektin I (MLI) erfolgte durch zwei unterschiedliche monoklonale Anti-MLI-Antikörper. Eine weitere Modellapplikation war die Visualisierung des nicht enzymatischen Schichtdickenabbaus des Polyethylencarbonats durch Superoxidradikalanionen unter Verwendung von murinen Makrophagen der Zelllinie J774A.1. Die Abbaugeschwindigkeit konnte mit den Triggerfaktoren Concanavalin A, LPS aus Escherichia coli und Salmonella typhimurium beeinflusst werden. Gram negative Bakterien als Trigger für Makrophagen wurden über ein Bakterienmembranmodell nachgeahmt. Hierfür wurden Liposomen mit Lipopolysaccharid aus Legionella pneumophila und Salmonella typhimurium funktionalisiert und mittels spezifischer Antikörper detektiert. Eine weitere Form des Triggerns der murinen Makrophagen der Zelllinie J774A.1 wurde mit LPS-gekoppelten Magnetic Beads auf der LAPS (lichtadressierbarer potentiometrischer Sensor) Messplattform umgesetzt. Unter Zuhilfenahme eines Permanentmagneten wurde ein Konzentrationsgradient in der Messlösung erzeugt. Die metabolische Aktivität der Makrophagen konnte so in Form der Änderung des pH-Wertes gezeigt werden.

Summary:
Using surface plasmon resonance (SPR) different model applications were established. Different sugar surfaces were prepared and characterised by a lectin screening. The binding behaviour of such surfaces was examined by NCI-H125 and Lewis Lung cells. In order to determine mistletoe lectin in drugs for cancer therapy a mistletoe lectin assay was developed. The detection of recombinant mistletoe lectin I (MLI) was performed by two different monoclonal anti-MLI-antibodies. Another model application was the visualisation of the non-enzymatic degradation of layer thickness of polyethylene carbonate by superoxide radical anions using murine macrophages of cell line J774A.1. The rate of degradation could be influenced by the trigger factors Concanavalin A, LPS from Escherichia coli and Salmonella typhimurium. Gram negative bacteria as a trigger for macrophages were imitated by a bacterial membrane model. For this purpose liposomes were functionalised with lipopolysaccharide from Legionella pneumophila and Salmonella typhimurium and detected using specific antibodies. Another mode of triggering murine macrophages from cell line J774A.1 was performed by the use of LPS-coupled magnetic beads implemented on the LAPS (light-addressable potentiometric sensor) measurement platform. By utilisation of a permanent magnet a concentration gradient was produced in the test solution. The metabolic activity of macrophages could be measured as a change in pH-value.

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