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. 2017 Jun 9;76(7):1176–1183. doi: 10.1136/annrheumdis-2016-210059

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Representations showing the development of antibody after immunisation of rabbits with carbamylated vimentin (carbVim). (A) Kinetics of antibody development after immunisation of rabbits with carbVim, vimentin and mutated citrullinated vimentin (MCV). Only immunisations with carbVim induced the formation of anti-carbVim antibodies. Human IgG-Fc from serum samples of healthy controls was isolated employing protein-A-affinity chromatography, separated under denatured conditions in SDS-PAGE and transferred onto nitrocellulose membrane for immunoblotting. IgG heavy chains were recognised by rabbit serum immunised with carbVim. (B) Differences in antibody specificities (profiles) after immunisation of rabbits with either carbVim (K1-3) or citrullinated vimentin (citrVim) (K4-6). Rabbits were immunised with 20 µg of the indicated antigen in Freund's complete adjuvant. Sera from the indicated animals were 1:200 diluted and analysed with the indicated antigens using enzyme immune assay (observed optical absorption are shown from a representative assay). RF, rheumatoid factor.