Analysis of pigeon intestinal mucin allergens using a novel dot blot assay

Many glycoproteins contain multiple glycosylation sites that can present mu lti-valent epitopes for antigenic recognition. Release of the oligosacchari des results in loss of avidity of antibody binding, which has been overcome by reforming clustered ligands, usually by reductive amination of free red ucing oligosaccharides to poly-amine groups. We have adapted this approach to hydrazinolytic release of O-linked chains of mucin glycoproteins and 'on e-pot' microscale coupling to poly-L-lysine (PLL). The conjugated PLL adher es to nitrocellulose membranes through washing procedures required for anti body or lectin overlay and detection. We show evidence for the applicabilit y of this technique using lectin and antibody reactivity to the oligosaccha rides of pigeon intestinal mucins, which have been implicated in the allerg ic disease pigeon fanciers' lung. (C) 2000 Elsevier Science Ltd. Ail rights reserved.