PMID: 8960101Nov 29, 1996Paper

Comparison of two different methods using overlapping synthetic peptides for localizing linear B cell epitopes in the U1 snRNP-C autoantigen

Journal of Immunological Methods
H HalimiSylviane Muller

Abstract

We have compared the performances of two different approaches using overlapping synthetic peptides to identify the location of linear epitopes of the U1 snRNP-C autoantigen. The first method was based on the use of 15 overlapping peptides (16-30 residue-long) synthesized using conventional Fmoc chemistry, removed from the resin by a standard cleavage procedure, and tested by ELISA after direct coating to polyvinyl microtiter plates. The second approach used a commercial kit (SPOT) to synthesize 75 overlapping decapeptides on cellulose membrane which were assayed by a direct immunoenzymatic test. Both standard and SPOTscan methods were evaluated with antibodies raised in rabbits against synthetic peptides of U1C and sera from patients with autoimmune diseases. In addition to inherent problems linked to the SPOT synthesis (in particular the impossibility of checking the quality of peptides), a number of limitations in the SPOTscan method were identified (e.g. a certain lack of sensitivity and, in one case, the complete lack of peptide reactivity due to the removal of charged end groups at both extremities). However, we found no background with sera from autoimmune patients in the SPOTscan and the antigenic maps obtained using the...Continue Reading

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Citations

Jan 22, 2004·Clinica Chimica Acta; International Journal of Clinical Chemistry·John G RoutsiasHaralampos M Moutsopoulos
May 24, 2003·Clinical Immunology : the Official Journal of the Clinical Immunology Society·M MahlerK M Pollard
Oct 26, 1999·The Journal of Peptide Research : Official Journal of the American Peptide Society·A KramerJ Schneider-Mergener
Apr 1, 2006·Critical Reviews in Clinical Laboratory Sciences·John G RoutsiasAthanasios G Tzioufas
Feb 28, 2009·Arthritis and Rheumatism·Brian D PooleJudith A James
Feb 22, 2011·Journal of Autoimmunity·E I KampylafkaA G Tzioufas
Dec 7, 2007·Biotechnology & Genetic Engineering Reviews·Kai HilperRobert E W Hancock
Apr 17, 2013·International Journal of Speech-language Pathology·Gillian SteelRosalind Thornton

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