The production of recombinant single chain antibody fragments for the detection of illicit drug residues
Brennan, Joanne
(2005)
The production of recombinant single chain antibody fragments for the detection of illicit drug residues.
PhD thesis, Dublin City University.
Recombinant antibodies represent a more sensitive and specific detection tool for immunoanalysis. The research carried out for this thesis describes the production of genetically-derived single chain antibody fragments to detect illicit drugs.
A variety of novel recombinant antibody fragments against morphine-3-glucuronide, a metabolite of heroin has been produced. A monomeric, dimeric and enzyme-labelled scFv were characterised with respect to their binding abilities and cross reactivities. Monomeric scFv was successfully applied to a competitive ELISA format for the detection of morphine residues in saliva. The assay was used to positively identify morphine residues in the saliva of drug addicts attending a rehabilitation clinic. An inhibition assay to detect morphine in saliva was also developed using the BIAcore 3000 instrument, a surface plasmon resonance-based biosensor for detection of biomolecular interactions in ‘real-time’. A novel sol particle lateral flow immunoassay was generated using monomeric and dimeric scFvs for the detection of M3G.
Recombinant antibody fragments against tetrahydrocannabinol were isolated from a naive human library by a process known as biopanning. Phage-displayed antibodies have been fully characterised with respect to their binding capabilities and were successfully applied to an immunoassay for the detection of marijuana residues in saliva.
Item Type:
Thesis (PhD)
Date of Award:
2005
Refereed:
No
Supervisor(s):
O'Kennedy, Richard
Uncontrolled Keywords:
illicit drug detection; heroin; recombinant antibodies; genetically-derived single chain antibody fragments