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'Real-time' biomolecular interaction analysis: novel applications and developments

Quinn, John G. (1999) 'Real-time' biomolecular interaction analysis: novel applications and developments. PhD thesis, Dublin City University.

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Abstract

The work described in this thesis involves the application of direct 'real-time' biomolecular interaction analysis using existing SPR-based biosensors and the development of novel alternative SPR-based technologies. These systems were applied to the investigation of whole cell binding to an immobilised ligand. Solid-phase blood typing was demonstrated by employing monoclonal anti- A IgG and anti-B IgM as affinity ligands for the capture of washed whole red blood cells (RBC). Protein A affinity-capture of anti-A IgG was used as an alternative to direct immobilisation thereby facilitating surface regeneration. A new non-'real-time' fibre optic-based system (BIACORE Probe) was evaluated using the same affinity-capture format and was successfully applied to direct whole blood typing. Non-'real time' analysis limited the data quality. Therefore, a similar 'real-time' fibre optic device was constructed. In addition, a miniature SPR-based biosensor employing a single platform optical configuration was developed with a view towards the production of a smaller, cheaper and portable alternative to the existing range of instruments. Determination of kinetic rate constants using direct 'real-time' biosensors has proved more complex than suggested by early investigators and has lead to misleading conclusions from artifactual data. A model study, employing the interaction of CD4 and glutathione-s-transferase with specific monoclonal antibodies, investigated experimental and data analysis methods for the determination of reliable rate constants. In addition, novel assays for determination of the active analyte concentration without the requirement for a standard curve were examined.

Item Type:Thesis (PhD)
Date of Award:1999
Refereed:No
Supervisor(s):O'Kennedy, Richard
Uncontrolled Keywords:Molecular recognition; Biomolecular interaction analysis
Subjects:Biological Sciences > Biotechnology
Biological Sciences > Cell biology
DCU Faculties and Centres:DCU Faculties and Schools > Faculty of Science and Health > School of Biotechnology
Use License:This item is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 License. View License
ID Code:19289
Deposited On:19 Sep 2013 12:24 by Celine Campbell. Last Modified 19 Sep 2013 12:24

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