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Peptide-based Identification of Functional Motifs and their Binding Partners
Published on: June 30, 2013
Random glycopeptide bead libraries for seromic biomarker discovery
Stjepan K Kracun1, Emiliano Cló, Henrik Clausen
1Department of Cellular and Molecular Medicine, University of Copenhagen, Copenhagen Center for Glycomics, Blegdamsvej 3b, DK-2200 Copenhagen N, Denmark.
Journal of Proteome Research
|October 5, 2010
Summary
We developed a novel glycopeptide bead library platform for discovering disease biomarkers. This high-throughput method enables rapid identification of autoantibodies and other binding proteins for early disease diagnosis and management.
Area of Science:
- Biochemistry
- Immunology
- Biotechnology
Background:
- Disease-specific biomarkers are crucial for early diagnosis and management.
- Aberrant protein post-translational modifications (PTMs), like O-glycosylations (O-PTMs), can trigger autoantibodies serving as sensitive biomarkers.
Purpose of the Study:
- To develop a random glycopeptide bead library screening platform for detecting autoantibodies and other binding proteins.
- To demonstrate the platform's utility in identifying tumor-specific glycopeptide biomarkers.
Main Methods:
- Construction of glycopeptide libraries on PEGA beads with a safety-catch C-terminal amide linker (SCAL).
- Mild cleavage conditions for glycopeptide release and sequence determination using ESI-Orbitrap-MS(n).
- On-bead enzymatic glycosylation to diversify glycoforms and microarray printing for validation.
Main Results:
- Successful detection of tumor-specific glycopeptide reporter epitopes using monoclonal antibodies and cancer patient autoantibodies.
- Resynthesis and validation of identified glycopeptides on microarrays.
- Diversification of O-glycopeptide libraries to various tumor glycoforms via enzymatic reactions.
Conclusions:
- A high-throughput, flexible platform for rapid discovery of O-glycopeptide biomarkers has been developed.
- The platform is applicable to lectin, antibody, and enzyme specificity studies, as well as other PTM investigations.
