• Libraries
    • Log in to:
    View Item 
    •   MSpace Home
    • Faculty of Graduate Studies (Electronic Theses and Practica)
    • FGS - Electronic Theses and Practica
    • View Item
    •   MSpace Home
    • Faculty of Graduate Studies (Electronic Theses and Practica)
    • FGS - Electronic Theses and Practica
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Bypassing immunization in an attempt to develop beta 1 specific monoclonal human antibodies from semi-synthetic repertoires

    Thumbnail
    View/Open
    MQ51687.pdf (5.109Mb)
    Date
    1999-06-01
    Author
    Binda, Chantal
    Metadata
    Show full item record
    Abstract
    Immune selection can be mimiced 'in vitro' by displaying antibody repertoires on the surface of a filamentous bacteriophage and selecting phage by binding to antigen. The development of recombinant DNA methods and the knowledge that antibody fragments could be functionally expressed and assembled in 'E. coli' have led to the development of phage-antibody display technology. Integrins are conserved proteins involved in tissue morphogenesis, tissue integrity, develop ent, inflammation, control of cell growth, and cell mobility. Highly conserved regions of the integrins are thought to play an important role in their function. Failure to raise antibodies against those regions using conventional hybridoma technology has led to the use of semi-synthetic repertoires in an attempt to develop B1 integrin specific monoclonal antibodies. Large diversity is an essential feature of semi-synthetic repertoires of antibodies. Seven semi-synthetic phage-antibody human libraries were used, where HCDR3 was randomised over 7 or 13 amino acid residues. These randomized heavy chains where cloned with a unique Humkv 325 light chain as Fab fragments into pComb 3H vector for phage display. The libraries were panned by binding to a biotinylated B1 integrin peptide known to be the epitope of an inhibitory antibody called JB1A, or to purified human B1 integrin. Seven rounds of panning, with increasing stringency, were done in an attempt to increase the proportion of highly specific clones. Modest enrichment was observed for the selection on peptide and no enrichment was observed on purified human B1 integrin. Clones selected using these semi-synthetic antibody libraries do not seem to have any specificity for the desired epitope. Improvement of the method of selection and system outlined in this thesis should be done in any future attempt of selection. Nevertheless phage-antibody display technology has a promising future as a complement to the well established hybridoma technology.
    URI
    http://hdl.handle.net/1993/2221
    Collections
    • FGS - Electronic Theses and Practica [25494]

    DSpace software copyright © 2002-2016  DuraSpace
    Contact Us | Send Feedback
    Theme by 
    Atmire NV
     

     

    Browse

    All of MSpaceCommunities & CollectionsBy Issue DateAuthorsTitlesSubjectsThis CollectionBy Issue DateAuthorsTitlesSubjects

    My Account

    Login

    Statistics

    View Usage Statistics

    DSpace software copyright © 2002-2016  DuraSpace
    Contact Us | Send Feedback
    Theme by 
    Atmire NV