Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.628614
Title: Cloning and analysis of thyroid stimulating hormone antibodies
Author: Qowaider, Salema
ISNI:       0000 0004 5346 5602
Awarding Body: University of Sheffield
Current Institution: University of Sheffield
Date of Award: 2014
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Abstract:
In this study we were able to synthesis a control TSAb monoclonal phage antibody construct and investigate binding of this and related reagents to the human TSH receptor. We are the first group to report stimulatory activity of a TSAb in scFv format, a finding that will contribute to our understanding of the mechanism of receptor stimulation by autoantibodies. We were able to demonstrate that the conformation of the native receptor rendered the target stimulatory epitope inaccessible to phage antibody constructs. This was an important finding that explained previous failed attempts, both reported and unreported, to enrich receptor antibodies by phage display. We went on to investigate this phenomenon at length and were finally able to resolve these difficulties by generating a novel cell line which expressed a receptor sub-domain (amino acids 1-260) anchored by a GPI motif. We were able to demonstrate that this receptor construct was able to specifically bind TSAb monoclonal antibody in phage form, a discovery that will open the door to future phage enrichment studies, and may contribute to the isolation of further examples of TSAb monoclonals from patient samples. Finally, we became the first research group to generate an anti-idiotypic monoclonal antibody that was able to bind and negate the antigen recognition site of the M22 TSAb antibody. This reagent should enable an entire series of novel experiments involving isolation and recovery of 'M22-like' antibodies and corresponding B cells from patients samples. In addition this anti-idiotypic reagent should offer a novel selective strategy for the recovery of stimulatory antibodies from recombinant antibody libraries.
Supervisor: Watson, Phil Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.628614  DOI: Not available
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