2013addispwphd.pdf (27.21 MB)
Download fileStructural Characterisation of Antibody/Antigen Interactions: Implications for B-Cell Receptor Signalling
thesis
posted on 2017-02-10, 13:45 authored by Philip William AddisTherapeutic antibodies are an important class of pharmaceutical molecules contributing
a significant portion of the total therapeutic market. Due to their high specificity,
modular nature and low toxicity, these proteins provide great potential for the
development of novel therapeutics for a wide range of diseases. The continued
development of improved and unique therapeutics will benefit from improvements in
our understanding of the characteristics of these proteins and their functions, facilitating
a more informed approach towards therapeutic research and potentially highlighting
new molecular information on poorly understood areas of antibody function, such as the
initiation of BCR signalling. NMR provides a useful and highly sensitive tool for the
investigation of the structural properties of proteins and the changes which occur upon
binding. Smaller antibody fragments also provide a useful model for the study of
antibody/antigen interactions in solution. The work described in this thesis describes the
development of the scFv format of antibody fragment as an amenable and reliable
model system from which high quality NMR data can be collected to investigate both
the behaviour of the antigen recognising CDR loops and the movements occurring at
the interface between the variable domains. An efficient, high yield protein expression
method was adapted and developed for the expression of large quantities of 15N/13C/2H
labelled scFv which allowed, for the first time, sequence specific backbone assignments
for the majority of the scFv in both the free and bound states and the comparison of
chemical shift data. Further to this, based upon the domain movements observed,
potential mechanisms for the initiation of BCR signalling were investigated resulting in
data suggesting the first antigen modulated molecular basis for the coupling of binding
from a structurally diverse antigen repertoire to a conserved signalling response.
History
Supervisor(s)
Carr, MarkDate of award
2013-02-01Author affiliation
Department of BiochemistryAwarding institution
University of LeicesterQualification level
- Doctoral
Qualification name
- PhD