PMID: 8946014Nov 13, 1996Paper

Mapping of epitopes recognized by PM/Scl autoantibodies with gene-fragment phage display libraries

Journal of Immunological Methods
M BlüthnerF A Bautz

Abstract

Sera from patients suffering from the polymyositis/scleroderma overlap syndrome (PM/Scl) recognize two antigenically non-related proteins with apparent molecular masses of 100 kDa and 75 kDa respectively. The two proteins are part of a particle termed PM/Scl localized in the granular component of the nucleolus. The predominant immunoreactivity of the PM/Scl sera was shown to be directed against the 100 kDa protein. The cDNA of the 100 kDa protein has been cloned recently and its immunogenic regions have been partially mapped using recombinant proteins. Thus far the localization of antigenic determinants on polypeptides has been done by expressing defined cDNA fragments in bacteria or by synthesizing overlapping short peptides and probing their immunoreactivity with antibodies. Here we present an alternative approach to localize autoimmune epitopes using sera containing polyclonal antibodies and gene-fragment phage display libraries. For epitope fine mapping of the PM/Scl-100 protein random fragments of the corresponding cDNA were cloned into the PIII protein of fUSE-5. These gene-fragment phage display libraries were incubated with affinity purified anti-PM/Scl-100 antibodies to enrich for epitope-displaying phages. All PM/Scl ...Continue Reading

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Citations

Jan 16, 1999·Clinical Rheumatology·S Jablonska, M Blaszczyk
Feb 14, 2004·Clinical Immunology : the Official Journal of the Clinical Immunology Society·Laura M StintonMarvin J Fritzler
Aug 7, 1997·Journal of Immunological Methods·F FackE K Bautz
May 7, 1999·Immunotechnology : an International Journal of Immunological Engineering·J Fehrsen, D H du Plessis
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May 3, 2003·Genome Research·Paola ZacchiAndrew R M Bradbury
Sep 13, 2001·Infection and Immunity·B P MullaneyJ D Marks
May 1, 2001·Journal of Clinical Pathology·E C JuryW J Morrow
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May 19, 2005·Arthritis Research & Therapy·Michael MahlerMarvin J Fritzler
Apr 23, 2008·Current Protocols in Neuroscience·A Bradbury
Feb 12, 2010·Annals of the New York Academy of Sciences·Michael Mahler, Marvin J Fritzler
Apr 8, 2010·Current Protocols in Neuroscience·Andrew R M Bradbury
Jul 18, 2018·Biosensors·Jingyi QiuTianfu Wu
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Aug 23, 2005·Archives of Biochemistry and Biophysics·Yi LiWilliam k Chan

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