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Generation of Monoclonal Antibodies against Human Prion Proteins in PrP0/0 Mice

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Abstract

Background

Prion diseases belong to a group of neurodegenerative disorders affecting humans and animals. The human diseases indude kuru, Creutzfeldt-Jakob disease (CJD), Gerstmann-Sträussler-Scheinker syndrome (GSS), and fatal familial insomnia (FFI). The pathogenic mechanisms of the prion diseases are not yet understood. Monoclonal antibodies provide valuable tools in the diagnosis, as well as in the basic research, of several diseases; however, monospecific antisera or monoclonal antibodies (mAbs) against human prion proteins were, until now, not available.

Materials and Methods

We have developed an immunization protocol based on nucleic acid injection into nontolerant PrP0/0 mice. DNA or RNA coding for different human prion proteins including the mutated sequences associated with CJD, GSS, and FFI were injected into muscle tissue. Mice were primarily inoculated with DNA plasmids encoding the prion protein (PRNP) gene and boosted either with DNA, RNA, or recombinant Semliki Forest Virus particles expressing PRNP. Hybridomas were then prepared.

Results

Different mAbs against human prion proteins were obtained, and their binding behavior was analyzed by peptide enzyme-linked immunosorbent assay. Western blot, immunofluorescence, and immunoprecipita-tion. Their cross-reactivity with prion protein from other species was also determined. Our mAbs are directed against four different linear epitopes and may also recognize discontinuous regions of the native prion protein.

Conclusions

These antibodies should allow us to address questions concerning the nature of the prion protein as well as the initiation and progression of prion diseases. Moreover, these mAbs can now be used for the diagnosis of prion diseases of humans and animals.

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Acknowledgments

We thank Professor Charles Weissmann (Zurich, Switzerland) for providing the PrP0/0 mice. Technical help in establishing the hybridoma fusions was provided by H. Peters and U. Friedrichs (Göttingen, Germany). P. Liljeström and P. Berglund (Stockholm, Sweden) kindly provided the pSFV1 and helper 2 plasmids. The skillful technical assistance of T. Jürgens is gratefully acknowledged. This project was supported by Grant 01KI9463 from the German BMBF.

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Authors and Affiliations

  1. Department of Virology and Immunology, German Primate Center (DPZ), Kellnerweg 4, 37077, Goettingen, Germany

    Susanne Krasemann, Gerhard Hunsmann & Walter Bodemer

  2. Federal Research Center for Virus Diseases of Animals, Tübingen, Germany

    Martin H. Groschup & Silke Harmeyer

Authors
  1. Susanne Krasemann
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  2. Martin H. Groschup
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  3. Silke Harmeyer
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  4. Gerhard Hunsmann
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  5. Walter Bodemer
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Krasemann, S., Groschup, M.H., Harmeyer, S. et al. Generation of Monoclonal Antibodies against Human Prion Proteins in PrP0/0 Mice. Mol Med 2, 725–734 (1996). https://doi.org/10.1007/BF03401656

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  • DOI: https://doi.org/10.1007/BF03401656

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