A high-throughput O-glycopeptide discovery platform for seromic profiling

TY - JOUR

T1 - A high-throughput O-glycopeptide discovery platform for seromic profiling

AU - Blixt, Ola

AU - Cló, Emiliano

AU - Nudelman, Aaron Samuel

AU - Sørensen, Kasper Kildegaard

AU - Clausen, Thomas

AU - Wandall, Hans H.

AU - Livingston, Philip O.

AU - Clausen, Henrik

AU - Jensen, Knud Jørgen

N1 - Keywords: Autoantibodies; Breast Neoplasms; Cancer Vaccines; Female; Glycopeptides; Glycosylation; Glycosyltransferases; Humans; Mucin-1; Peptides; Proteomics; Tumor Markers, Biological

PY - 2010/10/1

Y1 - 2010/10/1

N2 - 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.

AB - 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.

U2 - 10.1021/pr1005229

DO - 10.1021/pr1005229

M3 - Journal article

C2 - 20726594

SN - 1535-3893

VL - 9

SP - 5250

EP - 5261

JO - Journal of Proteome Research

JF - Journal of Proteome Research

IS - 10

ER -