Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.253776
Title: Clonal diversity of the immune response to insulin
Author: Mirza, Intisar Hussain
ISNI:       0000 0001 3410 9803
Awarding Body: University of Southampton
Current Institution: University of Southampton
Date of Award: 1987
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Abstract:
The clonal diversity of the provoked antibody immune response to human insulin was examined by the isolation of antibody secreting hybridomas from BALB/c mice. A wide spectrum of antibody binding polymorphisms was found with epitope sites being located on both conserved and variable regions of the insulin molecule and involving both sequential and noncontiguous amino acid residues. Diversity was found in the immune response to insulin not only with respect to epitope restriction but also with respect to antibody affinity. Thus, while some antibodies showed subresidue specificity and discriminated between the presence and absence of a single methyl group, others were heteroclitic and bound more strongly to insulin variants. The first part of the study established optimum immunisation schedules for the raising of monoclonal antibodies to insulin. It was found that fusion efficiencies of lymph node fusions after hind footpad immunisation were greater than those of conventional spleen cell fusions after dorsal subcutaneous immunisation and intraperitoneal boost. Fusion efficiencies correlated with specific antibody serum titres. A number of monoclonal antibodies gave high binding signals in enzyme-linked immunosorbent assay (ELISA) but low binding signals in liquid phase radiobinding assay (RBA). A solid phase absorption assay was therefore developed for characterising the binding site and relative affinity of these antibodies for insulin variants. Furthermore, the role of clonal restriction and affinity in the detection of insulin antibodies in liquid phase RBA and solid phase ELISA was studied, giving insights into the thermodynamic aspects of these assays. Monoclonal technology was also applied towards the isolation of monoclonal antibodies from an insulin autoimmune patient. IgM monoclonal antibodies binding to insulin were produced.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.253776  DOI: Not available
Keywords: Medicine
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