Diagnostic value of anti-GM1 ganglioside serology and validation of the INCAT-ELISA

Clonality of anti-GM1 IgM antibodies in multifocal motor neuropathy and the Guillain-Barré syndrome

“…Serum anti-GM1 IgM antibody titres and light-chain use were determined using the validated ELISA of the Inflammatory Neuropathy Cause and Treatment (INCAT) group,6 using the peroxidase-conjugated goat-antihuman IgM, goat-antihuman κ Ig (Sigma, A7164) and goat-antihuman λ Ig (Sigma, A5175). The concentration of anti-GM1 IgM light chains was expressed as the equivalent of standard dilution series of a monoclonal IgG1 κ and a monoclonal IgG1 λ fraction purified from the plasma of myeloma patients with a purity of at least 95% (Sigma I5154 and Sigma I5029).…”

Improving the detection of IgM antibodies against glycolipids complexes of GM1 and Galactocerebroside in Multifocal Motor Neuropathy using glycoarray and ELISA assays

“…Results were expressed in fluorescence intensities. ELISA was performed according to the standardized INCAT-ELISA technique (Willison et al, 1999;Kuijf et al, 2005). Sera with optical density above 0.1 at 1/100 dilution were titrated using a 5-fold dilution.…”

“…Anti-ganglioside antibodies against GM1 are detected in 40-60% of MMN sera (Pestronk et al, 1997;Kuijf et al, 2005) using traditional enzyme-linked immunosorbent assay (ELISA). Wide variations in assay performance have been reported (van Schaik et al, 1995), indicating that ELISA and related methodologies should ideally be standardized for consistency between different laboratories (Willison et al, 1999).…”

“…Wide variations in assay performance have been reported (van Schaik et al, 1995), indicating that ELISA and related methodologies should ideally be standardized for consistency between different laboratories (Willison et al, 1999). As they can also be found in up to 10% of ALS cases (Kuijf et al, 2005) and in normal and other disease controls, IgM isotype anti-GM1 antibodies are not specific for MMN. Testing antibodies against heteromeric complexes of GM1 and galactocerebroside (GalC) increases the sensitivity of antibody detection in MMN using either ELISA (Pestronk et al, 1997;Nobile-Orazio et al, 2010) or a modified dot-blot, herein termed glycoarray (Galban-Horcajo et al, 2013).…”

GM3, GM2 and GM1 mimics designed for biosensing: chemoenzymatic synthesis, target affinities and 900MHz NMR analysis

“…These drawbacks may partly explain the variation and limited sensitivity and specificity of the current ELISA to detect the antibodies. 24 Consequently, pure and structurally analogous synthetic mimics of gangliosides would have significant advantages over their parent structures. The formation of such mimics, and in general of compounds that are able to block glycan recognition does, however, still remain a major challenge.…”