Prion Protein-specific Antibodies That Detect Multiple TSE Agents with High Sensitivity

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2014 Mar 11

PMID 24608105

Citations 11

Authors

Sandra McCutcheon

Jan P M Langeveld

Boon Chin Tan

Andrew C Gill

Christopher de Wolf

Stuart Martin

Lorenzo Gonzalez

James Alibhai

A Richard Alejo Blanco

Lauren Campbell

Nora Hunter

E Fiona Houston

Affiliations

Soon will be listed here.

Abstract

This paper describes the generation, characterisation and potential applications of a panel of novel anti-prion protein monoclonal antibodies (mAbs). The mAbs were generated by immunising PRNP null mice, using a variety of regimes, with a truncated form of recombinant ovine prion protein spanning residues 94-233. Epitopes of specific antibodies were mapped using solid-phase Pepscan analysis and clustered to four distinct regions within the PrP molecule. We have demonstrated the utility of these antibodies by use of Western blotting and immunohistochemistry in tissues from a range of different species affected by transmissible spongiform encephalopathy (TSE). In comparative tests against extensively-used and widely-published, commercially available antibodies, similar or improved results can be obtained using these new mAbs, specifically in terms of sensitivity of detection. Since many of these antibodies recognise native PrPC, they could also be applied to a broad range of immunoassays such as flow cytometry, DELFIA analysis or immunoprecipitation. We are using these reagents to increase our understanding of TSE pathogenesis and for use in potential diagnostic screening assays.

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References

Tang Y, Gielbert A, Jacobs J, Baron T, Andreoletti O, Yokoyama T . All major prion types recognised by a multiplex immunofluorometric assay for disease screening and confirmation in sheep. J Immunol Methods. 2012; 380(1-2):30-9. DOI: 10.1016/j.jim.2012.03.004. View

Lee I, Westaway D, SMIT A, Wang K, Seto J, Chen L . Complete genomic sequence and analysis of the prion protein gene region from three mammalian species. Genome Res. 1998; 8(10):1022-37. DOI: 10.1101/gr.8.10.1022. View

Jeffrey M, Gonzalez L, Chong A, Foster J, Goldmann W, Hunter N . Ovine infection with the agents of scrapie (CH1641 isolate) and bovine spongiform encephalopathy: immunochemical similarities can be resolved by immunohistochemistry. J Comp Pathol. 2005; 134(1):17-29. DOI: 10.1016/j.jcpa.2005.06.005. View

Stanker L, Scotcher M, Lin A, McGarvey J, Prusiner S, Hnasko R . Novel epitopes identified by anti-PrP monoclonal antibodies produced following immunization of Prnp0/0 Balb/cJ mice with purified scrapie prions. Hybridoma (Larchmt). 2012; 31(5):314-24. PMC: 3482378. DOI: 10.1089/hyb.2012.0022. View

Lezmi S, Martin S, Simon S, Comoy E, Bencsik A, Deslys J . Comparative molecular analysis of the abnormal prion protein in field scrapie cases and experimental bovine spongiform encephalopathy in sheep by use of Western blotting and immunohistochemical methods. J Virol. 2004; 78(7):3654-62. PMC: 371064. DOI: 10.1128/jvi.78.7.3654-3662.2004. View

Mccutcheon S, Hunter N, Houston F . Use of a new immunoassay to measure PrP Sc levels in scrapie-infected sheep brains reveals PrP genotype-specific differences. J Immunol Methods. 2005; 298(1-2):119-28. DOI: 10.1016/j.jim.2005.01.012. View

Paramithiotis E, Pinard M, Lawton T, LaBoissiere S, Leathers V, Zou W . A prion protein epitope selective for the pathologically misfolded conformation. Nat Med. 2003; 9(7):893-9. DOI: 10.1038/nm883. View

Yokoyama T, Kimura K, Tagawa Y, Yuasa N . Preparation and characterization of antibodies against mouse prion protein (PrP) peptides. Clin Diagn Lab Immunol. 1995; 2(2):172-6. PMC: 170122. DOI: 10.1128/cdli.2.2.172-176.1995. View

Brown K, Mabbott N . Evidence of subclinical prion disease in aged mice following exposure to bovine spongiform encephalopathy. J Gen Virol. 2013; 95(Pt 1):231-243. DOI: 10.1099/vir.0.058958-0. View

10.

Panigaj M, Brouckova A, Glierova H, Dvorakova E, Simak J, Vostal J . Underestimation of the expression of cellular prion protein on human red blood cells. Transfusion. 2010; 51(5):1012-21. DOI: 10.1111/j.1537-2995.2010.02924.x. View

11.

Tan B, Blanco A, Houston E, Stewart P, Goldmann W, Gill A . Significant differences in incubation times in sheep infected with bovine spongiform encephalopathy result from variation at codon 141 in the PRNP gene. J Gen Virol. 2012; 93(Pt 12):2749-2756. DOI: 10.1099/vir.0.039008-0. View

12.

Rubenstein R, Kascsak R, Papini M, Kascsak R, Carp R, LaFauci G . Immune surveillance and antigen conformation determines humoral immune response to the prion protein immunogen. J Neurovirol. 1999; 5(4):401-13. DOI: 10.3109/13550289909029481. View

13.

Feraudet C, Morel N, Simon S, Volland H, Frobert Y, Creminon C . Screening of 145 anti-PrP monoclonal antibodies for their capacity to inhibit PrPSc replication in infected cells. J Biol Chem. 2004; 280(12):11247-58. DOI: 10.1074/jbc.M407006200. View

14.

Rhie A, Kirby L, Sayer N, Wellesley R, Disterer P, Sylvester I . Characterization of 2'-fluoro-RNA aptamers that bind preferentially to disease-associated conformations of prion protein and inhibit conversion. J Biol Chem. 2003; 278(41):39697-705. DOI: 10.1074/jbc.M305297200. View

15.

Andrievskaia O, McRae H, Elmgren C, Huang H, Balachandran A, Nielsen K . Generation of antibodies against bovine recombinant prion protein in various strains of mice. Clin Vaccine Immunol. 2006; 13(1):98-105. PMC: 1356621. DOI: 10.1128/CVI.13.1.98-105.2006. View

16.

Horiuchi M, Karino A, Furuoka H, Ishiguro N, Kimura K, Shinagawa M . Generation of monoclonal antibody that distinguishes PrPSc from PrPC and neutralizes prion infectivity. Virology. 2009; 394(2):200-7. DOI: 10.1016/j.virol.2009.08.025. View

17.

Campbell L, Gill A, McGovern G, Jalland C, Hopkins J, Tranulis M . The PrP(C) C1 fragment derived from the ovine A136R154R171PRNP allele is highly abundant in sheep brain and inhibits fibrillisation of full-length PrP(C) protein in vitro. Biochim Biophys Acta. 2013; 1832(6):826-36. DOI: 10.1016/j.bbadis.2013.02.020. View

18.

Slootstra J, Puijk W, Ligtvoet G, Langeveld J, Meloen R . Structural aspects of antibody-antigen interaction revealed through small random peptide libraries. Mol Divers. 1996; 1(2):87-96. PMC: 7088776. DOI: 10.1007/BF01721323. View

19.

Kascsak R, Rubenstein R, Merz P, Fersko R, Carp R, Wisniewski H . Mouse polyclonal and monoclonal antibody to scrapie-associated fibril proteins. J Virol. 1987; 61(12):3688-93. PMC: 255980. DOI: 10.1128/JVI.61.12.3688-3693.1987. View

20.

Kanyo Z, Pan K, Williamson R, Burton D, Prusiner S, Fletterick R . Antibody binding defines a structure for an epitope that participates in the PrPC-->PrPSc conformational change. J Mol Biol. 1999; 293(4):855-63. DOI: 10.1006/jmbi.1999.3193. View