Objective
The primary objective of this project is to define major structural parameters involved in promoting antibody-mediated neutralization of human cytomegalovirus (CMV) and to design human antibodies with perfected functions in this respect. Since this virus is a major health concern in immunocompromised individuals, such as neonatally infected children, transplantation patients and AIDS patients, such advances have important implication for the diagnosis and treatment of such disease. The project will furthermore yield information on how an in-vitro molecular evolution of an antibody specificity should proceed, using rational design and in-vitro selection, to provide an optimally efficient antibody, which might serve as a lead candidate in future passive immunotherapy studies.
The ultimate goal will be achieved by combining, in a multi-disciplinary (immunotechnology, virology, medical virology and structural biology) approach, recently developed
(i)technologies to obtain antibodies/antibody fragments by phage display selection
(ii) methodology designed to study the molecular interactions in three dimensions between the antigen and selected antibodies
(iii) methodology focusing on the biological function (virus neutralization) of the developed antibodies.
More specifically, we will create antibody gene libraries, for use in phage display of antibodies and select these in-vitro on a variety of recombinant CMV antigens exposing glycoprotein B-related epitopes. This antigen has been chosen since it is known to be the most important target for CMV-neutralizing antibodies.
Antibody/antigen interactions will subsequently be characterized by: (i)their ultra fine specificity at the molecular level by reactivity pattern analysis against e.g. mutated antigens and defined clinical isolates and isolates with different cell tropism;
(ii) determination of the molecular structures of antibody variable domains, antigens and antibody/antigen complexes using NMR;
(iii) their reaction rate kinetics; (iv) their biological (virus-neutralizing) activity.
These factors will together be assessed to, in subsequent development stratagies through an iterative process, modify the originally developed antibodies by rational design and selection stratagies to maximize their biological, CMV-neutralizing, activity. This will be achieved by targeting specific complementarity determining regions (CDR) of the antibody variable regions with e.g. "CDR shuffling" and light/heavy chain shuffling. Analysis of reactivity characteristics, including assessment of antigen-antibody structures, will permit in-vitro evolution of existing specific antibodies. These approaches will ensure that the resulting selected antibodies display perfected ultra fine specificity, clinical isolate recognition and reaction rate kinetics specifications ultimately resulting in optimal biological (CMV-neutralizing) properties. In addition to providing antibodies with optimal biological activity, this project will define the reaction parameters and structural interactions which are important for their biological activity and provide information and reagents by which antibody reactivity to the relevant epitopes can be efficiently assessed by immunological analysis.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques.
- natural sciencesbiological sciencesmicrobiologyvirology
- medical and health scienceshealth sciencesinfectious diseasesRNA virusesHIV
- medical and health sciencesbasic medicineimmunologyimmunotherapy
- medical and health sciencesclinical medicinetransplantation
- natural sciencesbiological sciencesmolecular biologymolecular evolution
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Call for proposal
Data not availableFunding Scheme
CSC - Cost-sharing contractsCoordinator
220 07 LUND
Sweden