Analysis of the Role of Type 1 Core O-Glycans in the Binding of Anti-MUC1 Antibodies by Cytofluorometry and Synthetic Peptide/Glycopeptide Binding Inhibition Studies

摘要: A panel of 56 MAbs submitted to the ISOBM TD-4 (MUC1) Workshop were analysed in two systems. These systems designed screen for peptide type 1 core O-glycan-related reactivities. Using synthetic MUC1 mucin-related peptides and glycopeptides, tested relative binding affinities O-glycan-substituted structures. studies utilized a competitive format with native human adenocarcinoma-derived mucin as solid phase. This system allows analysis glycoform subspecificity each MAb. The second approach taken parallel, MCF-7 (BrCa) OVCAR (OVCa) cell lines which grown presence or absence phenyl-N-acetylgalactosaminide (p-gal), blocker O-linked glycosylation. cells by FACS examine role these same glycan substitutions play regard either diagnostic therapeutic application MAbs. By there was consistent increased ‘epitope exposure’ peptide-specific p-gal. In addition, single MAb (TD-4 #150) is interpreted react O-glycan, probably Tn, TF STn specificity.