Binding patterns of DTR-specific antibodies reveal a glycosylation-conditioned tumor-specific epitope of the epithelial mucin (MUC1).
Glycosylation determines essential biological functions of epithelial mucins in health and disease. We report on the influence of glycosylation of the immunodominant DTR motif of MUC1 on its antigenicity. Sets of novel glycopeptides were synthesized that enabled us to examine sole and combined effects of peptide length (number of repeats) and O-glycosylation with GalNAc at the DTR motif on the binding patterns of 22 monoclonal antibodies recognizing this motif. In case of unglycosylated peptides almost all antibodies bound better to multiple MUC1 tandem repeats. Glycosylation at the DTR led to enhanced binding in 11 cases, whereas 10 antibodies were not influenced in binding, and one was inhibited. In nine of the former cases both length and DTR glycosylation were additive in their influence on antibody binding, suggesting that both effects are different. Improved binding to the glycosylated DTR motif was exclusively found with antibodies generated against tumor-derived MUC1. Based on these data a tumor-specific MUC1 epitope is defined comprising the ...PDTRP... sequence in a particular conformation essentially determined by O-glycosylation at its threonine with either GalNAcalpha1 or a related short glycan. The results can find application in the field of MUC1-based immunotherapy.
Karsten
Uwe
U
NEMOD GmbH & Co. KG, Robert-Rössle-Str. 10, D-13125 Berlin, Germany. uwe.karsten@nemod.com
Serttas
Nida
N
Paulsen
Hans
H
Danielczyk
Antje
A
Goletz
Steffen
S
D020816:Amino Acid Motifs; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000915:Antibody Affinity; D014408:Biomarkers, Tumor / Q000737:chemistry*; D004848:Epithelium / Q000737:chemistry; D018604:Epitope Mapping; D000939:Epitopes; D006020:Glycopeptides / Q000138:chemical synthesis / Q000737:chemistry; D006031:Glycosylation; D006801:Humans; D018396:Mucin-1 / Q000737:chemistry*; D010084:Oxidation-Reduction; D010446:Peptide Fragments; D010504:Periodic Acid; D020080:Tandem Repeat Sequences
D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D000939:Epitopes; D006020:Glycopeptides; D018396:Mucin-1; D010446:Peptide Fragments; D010504:Periodic Acid