A high-throughput O-glycopeptide discovery platform for seromic profiling.
Biomarker microarrays are becoming valuable tools for serological screening of disease-associated autoantibodies. Post-translational modifications (PTMs) such as glycosylation extend the range of protein function, and a variety of glycosylated proteins are known to be altered in disease progression. Here, we have developed a synthetic screening microarray platform for facile display of O-glycosylated peptides (O-PTMs). By introduction of a capping step during chemical solid-phase glycopeptide synthesis, selective enrichment of N-terminal glycopeptide end products was achieved on an amine-reactive hydrogel-coated microarray glass surface, allowing high-throughput display of large numbers of glycopeptides. Utilizing a repertoire of recombinant glycosyltransferases enabled further diversification of the array libraries in situ and display of a new level of potential biomarker candidates for serological screening. As proof-of-concept, we have demonstrated that MUC1 glycopeptides could be assembled and used to detect autoantibodies in vaccine-induced disease-free breast cancer patients and in patients with confirmed disease at time of diagnosis.
Blixt
Ola
O
Copenhagen Center for Glycomics, Department of Cellular & Molecular Medicine, University of Copenhagen, Blegdamsvej 3B, Copenhagen, Denmark. olablixt@sund.ku.dk
Cló
Emiliano
E
Nudelman
Aaron S
AS
Sørensen
Kasper Kildegaard
KK
Clausen
Thomas
T
Wandall
Hans H
HH
Livingston
Philip O
PO
Clausen
Henrik
H
Jensen
Knud J
KJ
D001323:Autoantibodies / Q000097:blood* / Q000276:immunology; D014408:Biomarkers, Tumor / Q000097:blood* / Q000276:immunology; D001943:Breast Neoplasms / Q000097:blood / Q000175:diagnosis / Q000276:immunology; D019496:Cancer Vaccines / Q000276:immunology; D005260:Female; D006020:Glycopeptides / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D006031:Glycosylation; D016695:Glycosyltransferases / Q000378:metabolism; D006801:Humans; D018396:Mucin-1 / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D010455:Peptides / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D040901:Proteomics / Q000379:methods*
D001323:Autoantibodies; D014408:Biomarkers, Tumor; D019496:Cancer Vaccines; D006020:Glycopeptides; C060192:MUC1 protein, human; D018396:Mucin-1; D010455:Peptides; D016695:Glycosyltransferases