The humanisation and generation of antibody fragments against oxidised low density lipoprotein for the optical molecular imaging of atherosclerosis
File(s)
Author(s)
Pandey, Samata Shrestha
Type
Thesis
Abstract
Atherosclerosis and its major adverse cardiovascular events remain a leading cause of mortality and morbidity globally. Despite advances in diagnostics and treatment, a key barrier to reducing its consequences is the difficulty in predicting and identifying those plaques which are most vulnerable to rupture. Monoclonal antibodies provide a unique opportunity in tackling this gap due to their natural high specificity for an epitope; making them promising imaging agents for the molecular imaging of atherosclerosis. In this effort, single chain variable fragments (scFv) and fragment antigen binding (Fab) of a characterised murine monoclonal antibody, LO1, have been generated for the molecular imaging of oxidised low density lipoprotein. These small size of these antibody fragments may optimise tissue penetrance and kinetics for molecular imaging, generating increased signal to noise ratio and greater targeting. In a translational effort, a humanised Fab was generated to minimise potential immunogenicity. The rationale design and expression of the humanisation and antibody fragment generation is presented. Near-infrared labelling of the humanised Fab allowed non-invasive imaging of atherosclerosis in a mouse model using fluorescence molecular tomography (FMT). Results revealed the humanised Fab successfully distinguishes atherosclerotic animals from wild-type mice. Multiplexed imaging with fluorescent probes targeting matrix metalloproteinases and integrin α5β3 discovered the humanised Fab was able to distinguish between diseased and non-diseased animals with the greatest confidence. Overall, this thesis demonstrates the utility of antibody fragments for molecular imaging.
Version
Open Access
Date Issued
2021-03
Date Awarded
2021-10
Copyright Statement
Creative Commons Attribution NonCommercial NoDerivatives Licence
Advisor
Kontoravdi, Kleio
Haskard, Dorian
Sternberg, Michael
Sponsor
British Heart Foundation
Publisher Department
Department of Chemical Engineering
Publisher Institution
Imperial College London
Qualification Level
Doctoral
Qualification Name
Doctor of Philosophy (PhD)