McDonnell, Barry (2010) Development of novel antibody-based diagnostics for the early and rapid detection of cardiac markers. PhD thesis, Dublin City University.
Abstract
The principal objective of this research was the production, characterisation and application
of antibodies for the detection of both C-reactive protein (CRP) and Myeloperoxidase
(MPO), two promising candidates in the field of cardiac diagnostics. CRP is a highly stable
acute phase reactant, produced in the liver in response to interleukin (IL)-6. In January 2003
both the Centres for Disease Control and Prevention (CDC) and the American Heart
Association (AHA) recommended CRP as the inflammatory marker of choice for assessment
of cardiovascular risk. The other biomarker under investigation was MPO, a mediator
enzyme secreted by inflammatory cells such as activated neutrophils and monocytes. High
levels of MPO have been reported in atherosclerotic plaques and accumulating evidence
indicates that MPO may have a causative role in plaque vulnerability. It has been suggested
that these markers may even be complementary in the assessment of patient cardiac risk. As a
marker of disease activity and vascular inflammation, CRP detection is associated with longterm risk stratification while MPO, being a marker of plaque instability and neutrophil
activation, may be prove to be a key marker for short term stratification.
A key feature of this work was to ensure the isolated antibodies had a sufficient level of both
functional affinity and specificity for the development of novel diagnostic formats and assay
platforms. Both classic polyclonal antibody generation and ‘state-of-the-art’ phage display
technology were employed for the generation of CRP- and MPO-specific antibodies and
recombinant fragments. The immune systems of mice, rabbits and chickens were investigated
in this work, which allowed for the identification of the main advantages and disadvantages
associated with each of the host immune systems and their capacity for generating specific
and sensitive antibodies. It was observed that the avian immune system was found to be the
most effective system for the generation of functional and soluble antibodies.
The CRP-specific antibodies were incorporated into three novel biosensor platforms for their
final characterisation. In addition, a novel lateral flow immunoassay (LFIA) for the rapid
detection of MPO was developed, using a recombinant antibody isolated from a MPOimmune library. Spiked samples of depleted human serum were employed in both the
biosensor and LFIA systems, thus validating the detection of the analytes in a realistic sample
matrix
Metadata
Item Type: | Thesis (PhD) |
---|---|
Date of Award: | 21 September 2010 |
Refereed: | No |
Supervisor(s): | O'Kennedy, Richard |
Uncontrolled Keywords: | Cardiology; C-reactive protein (CRP); Myeloperoxidase (MPO); Cardiac diagnostics |
Subjects: | Medical Sciences > Health |
DCU Faculties and Centres: | DCU Faculties and Schools > Faculty of Science and Health > School of Biotechnology Research Initiatives and Centres > Biomedical Diagnostics Institute (BDI) |
Use License: | This item is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 License. View License |
ID Code: | 15710 |
Deposited On: | 04 Apr 2011 14:00 by Dermot Walls . Last Modified 30 Jul 2021 15:02 |
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