Production and characterisation of a panel of monoclonal antibodies against native human cellular prion protein

Jones, Michael and McLoughlin, Victoria and Connolly, J.G. and Farquhar, Christine F. and MacGregor, Ian R. and Head, Mark W. (2009) Production and characterisation of a panel of monoclonal antibodies against native human cellular prion protein. Hybridoma, 28 (1). pp. 13-20. ISSN 1554-0014

Abstract

The human prion diseases, such as variant Creutzfeldt-Jakob disease (vCJD), are characterized by the conversion of the normal cellular prion protein (PrPC) into an abnormal disease associated form (PrPSc). Monoclonal antibodies (MAbs) that recognize these different PrP isoforms are valuable reagents both in the diagnosis of these diseases and in prion disease research in general but we know of no attempts to raise MAbs against native human PrPC. We immunized prion protein gene ablated (PrP-/-) mice with native human PrPC purified from platelets (pHuPrP) generating a predominantly IgG isotype anti-pHuPrP polyclonal antibody response in all mice. Following fusion of splenocytes from the immunized mice with SP2/0 myeloma cells, we were able to identify single cell clone and cryopreserve 14 stable hybridoma cell lines producing MAbs that reacted with pHuPrP. The properties of these MAbs (such as isotype, binding to native/denatured pHuPrP, and HuPrP epitopes recognized) are described. Furthermore, several of these MAbs showed a selectivity in their ability to immunoprecipitate disease associated PrPSc and its corresponding protease resistant core (PrPres).

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