A High-Throughput O-Glycopeptide Discovery Platform for Seromic Profiling

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• 作者：Ola Blixt ; Emiliano Cl ; Aaron S. Nudelman ; Kasper Kildegaard Sø ; rensen ; Thomas Clausen ; Hans H. Wandall ; Philip O. Livingston ; Henrik Clausen ; Knud J. Jensen
• 刊名：Journal of Proteome Research
• 出版年：2010
• 出版时间：October 1, 2010
• 年：2010
• 卷：9
• 期：10
• 页码：5250-5261
• 全文大小：758K
• 年卷期：v.9,no.10(October 1, 2010)
• ISSN：1535-3907

文摘

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.

<h4>Keywords:h4> Glycopeptide; synthesis; chemoenzymatic; enzyme; microarray; glycan array; autoantibodies; post-translational modification (PTM)