A straightforward protocol for the preparation of high performance microarray displaying synthetic MUC1 glycopeptides

摘要: Human serum MUC1 peptide fragments bearing aberrant O-glycans are secreted from columnar epithelial cell surfaces and known as clinically important biomarkers for the carcinoma when a specific monoclonal antibody can probe disease-relevant epitopes. Despite growing importance of glycopeptides biomarkers, precise epitopes most anti-MUC1 antibodies remains unclear.A novel protocol fabrication versatile microarray displaying peptide/glycopeptide library was investigated construction highly sensitive accurate epitope mapping assay various antibodies.Selective imine-coupling between aminooxy-functionalized methacrylic copolymer with phosphorylcholine unit synthetic glycopeptides-capped by ketone linker at N-terminus provided facile seamless preparation platform. It demonstrated that anti-KL-6 shows an extremely strong binding affinity toward carrying sialyl T antigen (Neu5Acα2,3Galβ1,3GalNAcα1→) Pro-Asp-Thr-Arg motif compared other seven such VU-3D1, VU-12E1, VU-11E2, Ma552, VU-3C6, SM3, DF3. The present also uncovered occurrence IgG autoantibodies in healthy human sera bind specifically attached five potential O-glycosylation sites tandem repeats.We established straightforward strategy standardized platform allowing analysis reducing background noise due to nonspecific protein adsorption.The approach would greatly accelerate discovery research new class well development therapeutic mAbs reacting