title: Human recombinant domain antibodies against multiple sclerosis antigenic peptide CSF114(Glc).
abstract: Multiple sclerosis (MS) is a chronic auto-immune disease characterized by a damage to the myelin component of the central nervous system. Self-antigens created by aberrant glycosylation have been described to be a key component in the formation of auto-antibodies. CSF114(Glc) is a synthetic glucopeptide detecting in vitro MS-specific auto-antibodies, and it is actively used in diagnostics and research to monitor and quantify MS-associated Ig levels. We reasoned that antibodies raised against this probe could have been relevant for MS. We therefore screened a human Domain Antibody library against CSF114(Glc) using magnetic separation as a panning method. We obtained and described several clones, and the one with the highest signals was produced as a 6×His-tagged protein to properly study the binding properties as a soluble antibody. By surface plasmon resonance measurements, we evidenced that our clone recognized CSF114(Glc) with high affinity and specific for the glucosylated peptide. Kinetic parameters of peptide-clone interaction were calculated obtaining a value of KD in the nanomolar range. Harboring a human framework, this antibody should be very well tolerated by human immune system and may represent a valuable tool for MS diagnosis and therapy, paving the way to new research strategies.
authors.affiliation: Dipartimento di Scienze Biomediche Sperimentali e Cliniche, Università degli Studi di Firenze, viale Morgagni 50, 50134, Firenze, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Niccheri
authors.forename: Feliciana
authors.initials: F
authors.lastname: Real-Fernàndez
authors.forename: Matteo
authors.initials: M
authors.lastname: Ramazzotti
authors.forename: Francesco
authors.initials: F
authors.lastname: Lolli
authors.forename: Giada
authors.initials: G
authors.lastname: Rossi
authors.forename: Paolo
authors.initials: P
authors.lastname: Rovero
authors.forename: Donatella
authors.initials: D
authors.lastname: Degl'Innocenti
mesh_terms: D000595:Amino Acid Sequence; D000906:Antibodies / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009103:Multiple Sclerosis / Q000175:diagnosis / Q000276:immunology*; D010455:Peptides / Q000737:chemistry / Q000276:immunology; D011994:Recombinant Proteins / Q000737:chemistry* / Q000276:immunology; D020539:Sequence Analysis, Protein; D020349:Surface Plasmon Resonance
chemical_list: D000906:Antibodies; D010455:Peptides; D011994:Recombinant Proteins
keywords: domain antibody; magnetic biopanning; multiple sclerosis