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.

Production of Antigen-Binding Fragment against O,O-Diethyl Organophosphorus Pesticides and Molecular Dynamics Simulations of Antibody Recognition

“…The 3D homology modeling structure of Fab was built via the SWISS-MODEL [ 32 ] and is shown in Figure 3 . Its application has been achieved for antibody homologous modeling [ 33 , 34 , 35 ]. As shown in Figure 3 A, Fab was composed by a heavy chain (V H -C H 1; Fd fragment) and a light chain (V L -C L , κ chain).…”

Expression, purification and characterization of anti-FGF7 domain antibody identified using phage display technique

“…Briefly, D53 coding genes located in phage pR2 genome were amplified and inserted into pGEX-6P-1 vector. However, DNA sequencing revealed the presence of an amber stop codon in the CDR2 region of D53, which is a problem frequently observed in phage display libraries 34,35 . The problem was resolved using site directed point mutation using overlap extension PCR mutagenesis technique to convert TAG amber stop codon into CAG to code for amino acid glutamine.…”

Phage Displayed Domain Antibodies (dAb) for Detection of Allergenic Pistachio Proteins in Foods

“…Moreover, a stop codon was present in the CDR1 of six clones, and in CDR3 or one clone, due to an amber mutation (Figure 4). This is frequent in phage displayed antibodies because of randomization of CDR sequences and propagation of phagemids in the amber suppressor TG1 E. coli strain [27]. Amber stop codons are interpreted as Gln residues in suppressor strains like TG1, so that these dAb clones can be used in a phage display Deduced amino acid sequences of the pistachio-binding clones showing the CDRs and immunoglobulin framework regions (FRs) demonstrated no insertions or deletions in CDR1 and CDR2 in any of the clones.…”