Novel approaches to target infectious diseases: the utility of Auger electrons and scFvs for imaging and control

Rathje, Claudia Cattoni (2014) Novel approaches to target infectious diseases: the utility of Auger electrons and scFvs for imaging and control. Doctor of Philosophy (PhD) thesis, University of Kent,.

Abstract

With an increasing number of infectious agents resistant to one or more existing antibiotics, there is a global requirement for new therapeutics. One possible method for treatment of infectious diseases proposed in this study is the adaptation of a technique developed for cancer treatment, targeted radionuclide therapy, with particular interest in Auger electron radiation. Auger electrons are short range and low energy particles emitted from a range of radioactive isotopes. As a targeting agent, the antimicrobial peptide fragment UBI 29-41 was used. Antimicrobial peptides have a broad range of target microorganisms, ideal for toxicological studies of Auger electrons. Here, the baseline toxicity of UBI 29-41 was established in a range of model organisms. This enabled the analysis of the toxic effect of Auger electron emitting radio-nuclides using the isotope125I. Although no direct toxic effect was observed with this particular isotope, the labelling allows easy replacement with more potent isotopes. To further develop the targeting agent a lipoarabinomannan (LAM) specific single-chain variable fragment (scFv) was developed. LAM is a glycolipid and virulence factor associated with M. tuberculosis, the primary cause of tuberculosis in humans. By isolating the variable domains of a LAM specific monoclonal antibody, linked via a poly-glycine linker, the scFv was assembled in silico. The nucleotide sequence was optimised for and transfected into a Pichia pink expression system. Expression was successful with a ~27kDa product being secreted. The novel anti-LAM scFv generated in this study combined with the potential toxic effects of Auger electrons could provide a new avenue for the treatment and diagnosis of tuberculosis.

Item Type:	Thesis (Doctor of Philosophy (PhD))
Thesis advisor:	Lloyd, Dan
Uncontrolled keywords:	Auger electrons, antibiotic, scFv, antimicrobial peptides, radiation, tuberculosis, protein expression, scFv design, Pichia pink, bacteria, yeast
Subjects:	Q Science > QP Physiology (Living systems) > QP517 Biochemistry
Divisions:	Faculties > Sciences > School of Biosciences
Depositing User:	Users 1 not found.
Date Deposited:	27 Feb 2015 12:12 UTC
Last Modified:	29 May 2019 14:16 UTC
Resource URI:	https://kar.kent.ac.uk/id/eprint/47432 (The current URI for this page, for reference purposes)