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
References
Jun 1, 1992·Molecular Biology Reports·I Pettersson
Jun 1, 1992·Molecular Biology Reports·K B Elkon
Jul 1, 1991·Immunology Today·A C HorsfallM G Jones
Feb 11, 1991·Nucleic Acids Research·R L NelissenW J van Venrooij
Jan 5, 1990·Science·V HeinrichsR Lührmann
Jun 1, 1986·Molecular Immunology·S MullerM H Van Regenmortel
Sep 24, 1987·Journal of Immunological Methods·H M GeysenP G Schoofs
Sep 12, 1988·Nucleic Acids Research·P T SillekensW J van Venrooij
Jul 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·H M GeysenS J Barteling
Mar 1, 1994·Molecular Immunology·N TuaillonS Muller
Jun 17, 1994·Cell·P A Sharp
Jun 1, 1994·Scandinavian Journal of Immunology·J A JamesJ B Harley
Aug 1, 1995·Proceedings of the National Academy of Sciences of the United States of America·T MimoriM Akizuki
Sep 1, 1995·The Journal of Biological Chemistry·J P BriandS Muller
Mar 1, 1993·Immunology Today·F C HayA Horsfall
May 1, 1993·European Journal of Immunology·Y MisakiK Ito
Oct 1, 1994·Molecular Immunology·C StemmerS Muller
Jan 1, 1996·Scandinavian Journal of Immunology·L R HaaheimR Jonsson
Citations
May 20, 1998·Arthritis and Rheumatism·P DeckerS Muller
Jan 22, 2004·Clinica Chimica Acta; International Journal of Clinical Chemistry·John G RoutsiasHaralampos M Moutsopoulos
Sep 29, 1999·Vaccine·F BrownS Muller
Oct 3, 1999·Vaccine·C D PartidosM W Steward
May 24, 2003·Clinical Immunology : the Official Journal of the Clinical Immunology Society·M MahlerK M Pollard
Nov 25, 1998·Nucleic Acids Research·H DumortierS Muller
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
Mar 3, 2010·Immunological Reviews·Nicole H KattahPaul J Utz
Oct 6, 2005·Biochimica Et Biophysica Acta·Hubert HalimiAlain De Caro
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