Histone Protein Epitope Mapping for Autoantibody Recognition in Rheumatoid Arthritis.

Feliciana Real-Fernández, Federico Pratesi, Paola Migliorini, Paolo Rovero

Journal: Methods in molecular biology (Clifton, N.J.) 2019;1901():221-228

PMID: 30539581

Abstract

Deiminated proteins are the target of diagnostic anti-citrullinated peptide/protein autoantibodies (ACPA) in rheumatoid arthritis (RA). Deiminated histone H4 contained in the neutrophil extracellular traps reacts with ACPA, becoming an interesting diagnostic antigen for RA. The identification of the ACPA binding site in histone H4 was performed experimentally by mapping the complete sequence. The method describes the synthesis of an overlapping peptide library covering the entire deiminated sequence of H4 and its further evaluation in ELISA. A detailed description of an ELISA protocol to test RA patients' sera against the synthesized peptides and ACPA is provided.

Address: Laboratory of Peptide and Protein Chemistry and Biology, Division of Pharmaceutical Sciences and Nutraceutics, Department of Neurofarba, University of Florence, Sesto Fiorentino, Italy.; Clinical Immunology and Allergy Unit, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy.; Clinical Immunology and Allergy Unit, Department of Clinical and Experimental Medicine, University of Pisa, Pisa, Italy. [email protected].; Laboratory of Peptide and Protein Chemistry and Biology, Division of Pharmaceutical Sciences and Nutraceutics, Department of Neurofarba, University of Florence, Sesto Fiorentino, Italy. [email protected].

DOI: 10.1007/978-1-4939-8949-2_18

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https://doi.org/10.1007/978-1-4939-8949-2_18

MeSH Terms: Amino Acid Sequence, Arthritis, Rheumatoid, Autoantibodies, Enzyme-Linked Immunosorbent Assay, Epitope Mapping, Histones, Humans, Immunoglobulin G, Peptide Library, Peptides, Solid-Phase Synthesis Techniques