Biomarker: N-glucosylation in Multiple Sclerosis

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Biomarker: N-glucosylation in Multiple Sclerosis

Post by frodo » Tue Jul 21, 2020 1:54 am

A Multiple N-Glucosylated Peptide Epitope Efficiently Detecting Antibodies in Multiple Sclerosis


Diagnostics of Multiple Sclerosis (MS) are essentially based on the gold standard magnetic resonance imaging. Few alternative simple assays are available to follow up disease activity. Considering that the disease can remain elusive for years, identification of antibodies fluctuating in biological fluids as relevant biomarkers of immune response is a challenge.

In previous studies, we reported that anti-N-glucosylated (N-Glc) peptide antibodies that can be easily detected in Solid-Phase Enzyme-Linked ImmunoSorbent Assays (SP-ELISA) on MS patients’ sera preferentially recognize hyperglucosylated adhesin of non-typeable Haemophilus Influenzae. Since multivalency can be useful for diagnostic purposes to allow an efficient coating in ELISA, we report herein the development of a collection of Multiple N-glucosylated Peptide Epitopes (N-Glc MEPs) to detect anti-N-Glc antibodies in MS. To this aim, a series of N-Glc peptide antigens to be represented in the N-GlcMEPs were tested in competitive ELISA. We confirmed that the epitope recognized by antibodies shall contain at least 5-mer sequences including the fundamental N-Glc moiety.

Using a 4-branched dendrimeric lysine scaffold, we selected the N-Glc MEP 24, carrying the minimal epitope Asn(Glc) anchored to a polyethylene glycol-based spacer (PEG) containing a 19-atoms chain, as an efficient multivalent probe to reveal specific and high affinity anti-N-Glc antibodies in MS.


The present study reported the development of new N-glucosylated Multivalent Epitope Peptides for the detection of anti-N(Glc) antibodies in Multiple Sclerosis. After a preliminary screening of several short linear peptide epitopes and multiple epitope peptides, the promising results suggest that multivalent interaction can be useful in designing SP-ELISA to detect antibodies to aberrant N-glucosylation in Multiple Sclerosis for both diagnostic and prognostic purposes. Indeed, the multivalent N-Glc MEP 24 ligand, carrying the minimal glucosylated epitope Asn (Glc) anchored to the 19-atoms PEG-based spacer, can be an efficient probe to reveal both IgM and IgG autoantibodies as disease biomarkers.

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