Di Pisa M.,University of Florence |
Buccato P.,DIESSE Ricerche Srl |
Sabatino G.,University of Florence |
Real Fernandez F.,Toscana Biomarkers SRL |
And 4 more authors.
Journal of Peptide Science | Year: 2014
Celiac disease (CD) is an autoimmune mediated disease with complex and multifactorial etiology. Gluten intake triggers a composite immune response involving T-cells and B-cells and leading to the secretion of autoantibodies if a genetic predisposition is present. Untreated CD patients show high levels of circulating autoantibodies directed to different auto-antigens present in the intestinal mucosa. The most important auto-antigen is the endomysial enzyme tissue transglutaminase (tTG). Both IgA and IgG antibody isotypes to tTG are known, but only the IgA antibodies demonstrate the highest disease specificity and thus are considered disease biomarkers. Because the pathogenicity and exact tTG binding properties of these autoantibodies are still unclear, the characterization of tTG antigenic domains is a crucial step in understanding CD onset and the autoimmune pathogenesis. Overlapping peptide libraries can be used for epitope mapping of selected protein portions to determine antigenic fragments contributing to the immunological activity and possibly develop innovative peptide-based tools with high specificity and sensitivity for CD. We performed an epitope mapping study to characterize putative linear auto-antigenic epitopes present in the tTG N-terminal portion (1-230). A library of 23 overlapping peptides spanning tTG(1-230) was generated by Fmoc/tBu solid-phase peptide synthesis and screened by immunoenzymatic assays employing patients' sera. The results indicate that four synthetic peptides, that is, Ac-tTG(1-15)-NH2, Ac-tTG(41-55)-NH2, Ac-tTG(51-65)-NH 2, and Ac-tTG(151-165)-NH2, are recognized by IgA autoantibodies circulating in CD patients' sera. These results offer important insight on the nature of the antigen-antibody interaction. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd.
Pandey S.,University of Florence |
Dioni I.,University of Florence |
Lambardi D.,University of Florence |
Real-Fernandez F.,University of Florence |
And 9 more authors.
Molecular and Cellular Proteomics | Year: 2013
Sophisticated approaches have recently led to the identification of novel autoantigens associated with Multiple Sclerosis (MuS), e.g. neurofascin, contactin, CNPase, and other T-cell receptor membrane anchored proteins. These putative antigens, although differing from the conventional myelin derivatives, are conceptually based on an animal model of experimental autoimmune encephalomyelitis. In this report we describe the identification of putative antigens based on their recognition by autoantibodies isolated from MuS patient serum. In a previous work from this laboratory we have shown that a peptide probe, named CSF114(Glc), specifically identifies serum autoantibodies in a subset of MuS patients, representing ⋯ 30% of the patient population. The autoantibodies, purified from MuS patients' sera (six), through CSF114(Glc) affinity chromatography, detected three immunoreactive protein bands present in the rat brain. Proteomic analysis of the immunoreactive bands, involving MALDI and MS/MS techniques, revealed the presence of four proteins distinguishable by their mass: alpha fodrin, alpha actinin 1, creatine kinase, and CNPase. The immunoreactive profile of these rat brain proteins was compared with that of commercially available standard proteins by challenging against either CSF114(Glc) purified MuS autoantibodies, or monoclonal antibodies. Further discrimination among the rat brain proteins was provided by the following procedure: whereas monoclonal antibodies recognized all rat brain proteins, isolated MuS specific antibodies recognize only alpha actinin 1 as a putative antigen. In fact, alpha actinin 1 displayed a robust immunoreactive response against all MuS patients' sera examined, whereas the other three bands were not consistently detectable. Thus, alpha actinin 1, a cyto-skeleton protein implicated in inflammatory/degenerative autoimmune diseases (lupus nephritis and autoimmune hepatitis) might be regarded as a novel MuS autoantigen, perhaps a prototypic biomarker for the inflammatory/degenerative process typical of the disease. © 2013 by The American Society for Biochemistry and Molecular Biology, Inc.
Papini A.M.,University of Florence |
Konig E.,University of Florence |
Konig E.,Toscana Biomarkers SRL
Expert Opinion on Therapeutic Patents | Year: 2015
Introduction: With > 2 million people affected by multiple sclerosis (MS) worldwide, the elucidation of its etiopathogenesis is of highest interest. Ongoing research in medicine, molecular biology, chemistry and physics aims to improve the life of MS patients by increasing efficacy and decreasing adverse side effects of presently available drugs. A precise diagnosis of this complex disease, which can take different courses, is fundamental to finding an efficient treatment strategy.Areas covered: We present a summary of diagnostic and therapeutic patents granted between 2009 and 2014. Diagnostic inventions use both genetic and proteomic approaches or measure cerebral venous hemodynamics. Instead, new treatments rely on small molecules and/or the active manipulation of proteins that are involved in the pathogenesis of MS.Expert opinion: There are some promising approaches among recently published patents. In particular, genetic profiling for diagnosis, combination of novel drugs with FDA-approved drugs to reduce side effects, and the personalisation of MS treatments according to a more defined diagnosis are considered as important. In the light of the latest developments, we discuss the complex picture of MS, which we assume to be different events connected by a causal chain consisting of circulatory abnormalities, altered redox processes in CNS immune cells, oligodendropathy, inflammation and finally autoimmunity. © Informa UK, Ltd.
Toscana Biomarkers S.R.L. | Date: 2012-05-24
The present invention refers to the use of citrullinated synthetic peptides for the diagnosis of rheumatoid arthritis (RA).
Toscana Biomarkers S.R.L. | Date: 2010-07-23
The present invention refers peptides and methods for the identification of diagnostic antibodies in auto-immune diseases such as Systemic Lupus Erythematosus (SLE), and therefore useful tools for diagnosis or therapeutic treatment of SLE.