Mass spectrometric and peptide chip epitope mapping of rheumatoid arthritis autoantigen RA33

European Journal of Mass Spectrometry
R F El-KasedM O Glocker

Abstract

The protein termed RA33 was determined to be one major autoantigen in rheumatoid arthritis (RA) patients and antiRA33 auto-antibodies were found to appear shortly after onset of RA. They are often detectable before a final diagnosis can be made in the clinic. The aim of our study is to characterise the epitope of a monoclonal antiRA33 antibody on recombinant RA33 using mass spectrometric epitope mapping. Recombinant RA33 has been subjected to BrCN cleavage and fragments were separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Subsequent in-gel proteolytic digestion and mass spectrometric analysis determined the partial sequences in the protein bands. Western blotting of SDS-PAGE-separated protein fragments revealed immuno-positive, i.e. epitope-containing bands. BrCN-derived RA33 fragments were also separated by high- performance liquid chromatography (HPLC) and immuno-reactivity of peptides was measured by dot-blot analysis with the individual HPLC fractions after partial amino acid sequences were determined. The epitope region identified herewith was compared to data from peptide chip analysis with 15-meric synthetic peptides attached to a glass surface. Results from all three analyses consiste...Continue Reading

References

Dec 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·D SuckauM Przybylski
Aug 21, 1986·Nature·D J BarlowJ M Thornton
Aug 19, 2000·Cancer Chemotherapy and Pharmacology·N SatoH Ikeda
Sep 12, 2002·Arthritis Research·Wolfgang HueberWilliam H Robinson
Dec 20, 2002·Zeitschrift für Rheumatologie·G Steiner, J S Smolen
Jun 13, 2003·Archives of Biochemistry and Biophysics·Aleksander CzogallaAleksander F Sikorski
Sep 5, 2003·Analytical and Bioanalytical Chemistry·Marcus MachtMichael Przybylski
Dec 30, 2004·Analytical Chemistry·Thomas KöcherRoman A Zubarev
Nov 12, 2005·Medicinal Research Reviews·Michael O GlockerHans-Jürgen Thiesen
Mar 11, 2006·Biomolecular Engineering·Marina CretichMarcella Chiari
Jun 6, 2006·The Journal of Immunology : Official Journal of the American Association of Immunologists·Christine Hager-BraunKenneth B Tomer
Jul 14, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·J C HeitnerM O Glocker
Mar 21, 2007·Zeitschrift für Rheumatologie·E FeistG-R Burmester
Sep 20, 2007·European Journal of Mass Spectrometry·Raluca StefanescuMichael Przybylski
Jan 5, 2008·Annual Review of Immunology·Lars KlareskogLars Alfredsson
Jan 25, 2008·BMC Bioinformatics·Yang Zhang
Apr 26, 2008·PLoS Computational Biology·Martin C FrithTimothy L Bailey
Feb 24, 2009·Proteomics·Christer WingrenCarl A K Borrebaeck
Mar 19, 2009·Proteomics·Oliver PoetzThomas O Joos
Apr 21, 2009·Methods in Molecular Biology·Suraj DhunganaKenneth B Tomer
Apr 21, 2009·Methods in Molecular Biology·Peter LorenzHans-Jürgen Thiesen
Oct 17, 2009·Expert Review of Neurotherapeutics·Gautam Prasad, Daphne A Haas-Kogan

❮ Previous
Next ❯

Citations

Jun 16, 2017·Journal of the American Society for Mass Spectrometry·Yelena YefremovaMichael O Glocker
Sep 14, 2018·Journal of Immunoassay & Immunochemistry·Reham F El-Kased
May 31, 2019·Molecular & Cellular Proteomics : MCP·Bright D DanquahMichael O Glocker
Aug 29, 2012·Molecular Biology International·Jaroslav KubrychtPavel Souček
Jul 13, 2016·Mass Spectrometry Reviews·Kwabena F M OpuniMichael O Glocker
Aug 1, 2019·Molecular & Cellular Proteomics : MCP·Bright D DanquahMichael O Glocker
Jan 7, 2015·Journal of the American Society for Mass Spectrometry·Yelena YefremovaMichael O Glocker

❮ Previous
Next ❯

Related Concepts

Related Feeds

Autoimmune Diseases

Autoimmune diseases occur as a result of an attack by the immune system on the body’s own tissues resulting in damage and dysfunction. There are different types of autoimmune diseases, in which there is a complex and unknown interaction between genetics and the environment. Discover the latest research on autoimmune diseases here.

Cancer Vaccines

Cancer vaccines are vaccines that either treat existing cancer or prevent development of a cancer.