Title |
Improved serological detection of rheumatoid arthritis: a highly antigenic mimotope of carbonic anhydrase III selected in a murine model by phage display
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Published in |
Arthritis Research & Therapy, June 2015
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DOI | 10.1186/s13075-015-0685-3 |
Pubmed ID | |
Authors |
Galber Rodrigues Araujo, Emília Rezende Vaz, Patricia Tiemi Fujimura, João Eurico Fonseca, Lucélia Maria de Lima, Helena Canhão, Gabriela Venturini, Karina Helena Morais Cardozo, Valdemir Melechco Carvalho, Marcelo Henrique Napimoga, Luiz Ricardo Goulart, João Gonçalves, Carlos Ueira-Vieira |
Abstract |
Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease that affects around 1 % of the human population worldwide. RA diagnosis can be difficult by the fact that there is no definitive test for its detection. Therefore, the aim of this study was to identify biomarkers that could be used for RA diagnosis. Sera from collagen-induced arthritis (CIA) mouse model were used to select potential biomarkers for RA diagnosis by Phage Display technology. In silico and in vitro analyses were performed to characterize and validate the selected peptides. Samples were classified into three groups: RA, two other immune-mediated rheumatic diseases (systemic lupus erythematosus - SLE and ankylosing spondylitis - AS) and healthy controls (HC). ELISA assay was carried out to determine antibody levels, and diagnostic parameters were determined by constructing Receiver Operating Characteristic (ROC) curves. Mass spectrometry and western blot were performed to identify the putative autoantigen that was mimicked by a highly reactive mimotope. After three rounds of selection, fourteen clones were obtained and tested for immunoreactivity analysis against sera from RA and HC groups. The phage-fused peptide with the highest immunoreactivity (M12) was synthesized, and was able of efficiently discriminate RA patients from SLE, AS and HCs (p < 0.0001) by ELISA. The specificity and sensitivity of anti-M12 antibodies for RA diagnosis were 91 % and 84.3 %, respectively. The M12 peptide was identified as a peptide that mimics a predicted antigenic site of the carbonic anhydrase III (CAIII) protein, a ubiquitous biomarker that has been identified in patients with other diseases. M12 is the first peptide associated to CAIII protein that may be used as an antigen for antibodies detection to aid in RA diagnosis with high sensitivity and specificity. |
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