MURAL - Maynooth University Research Archive Library

    Preparative separation of foreign antigens for highly efficient presentation to T cells in vitro

    Katrak, Kersi and Mahon, Bernard P. and Jones, Wendy C. and Briutigam, Sandra (1992) Preparative separation of foreign antigens for highly efficient presentation to T cells in vitro. Journal of Immunological Methods,, 156. pp. 247-254. ISSN 0022-1759

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    A method is described for the separation and purification of proteins from complex mixtures of foreign antigens in a form suitable for stimulating T cells in vitro. The technique involves electrophoretic separation of proteins followed by elution, concentration and adsorption of the polypeptide subunits to latex microspheres. Alternatively, where a specific antibody is available, proteins may be affinity-purified from a heterogeneous mixture of antigens, using antibody-coated latex microspheres. Nanogram quantities of protein coupled to latex were shown to be highly efficient stimulators of antigen-specific T cells as tested by in vitro proliferation and cytokine release assays. The utility of this technique was demonstrated using poliovirus capsid proteins separated by SDS-polyacrylamide gel electrophoresis (PAGE) and coupled to latex microspheres for specificity analysis of T cell clones. Antigen reactivity of the T cell clones was confirmed using recombinant baculoviruses expressing individual poliovirus proteins. Furthermore, recombinant proteins coupled to latex microspheres were used for efficient stimulation and in vitro propagation of T cell clones specific for the simian immunodeficiency virus (SIV) envelope (env) protein. Although the technique is illustrated in this report using viral antigens, it has also proved to be an efficient method for the separation of bacterial antigens in studies of polyclonal T cell responses to Bordetella pertussis antigens.

    Item Type: Article
    Keywords: Viral antigen; Bacterial antigen; T cell proliferation; T cell clone; Latex microsphere; SDS-PAGE;
    Academic Unit: Faculty of Science and Engineering > Biology
    Item ID: 7161
    Depositing User: Bernard Mahon
    Date Deposited: 06 Jul 2016 09:59
    Journal or Publication Title: Journal of Immunological Methods,
    Publisher: Elsevier
    Refereed: Yes
    Use Licence: This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available here

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