The development of high-throughput analytical techniques such as glycan microarrays has made dramatic changes in the field of glycomics. It provides a very valuable and time-and material-saving way to study protein-glycan interactions. However, in most of the cases, not only the glycan structure but also the presentation of the glycan structure on the peptide backbone is of importance in protein-glycan interactions. In addition, antibodies against a glycopeptide/glycoprotein usually recognize the peptide backbone as a part of the binding epitope. To this end, a novel glycopeptide microarray has been developed to screen antibodies raised against different glycopeptide antigens. This technique allowed identification of new binding epitopes and clearly revealed differences in the selectivity patterns for antibodies raised against different, yet closely related, glycopeptide antigens.
The glycopeptides are pre-synthesized accordingly and equipped with an N-terminal spacer, and then immobilized on N-hydroxysuccinimide ester (NHS) coated microarray slides. In some cases, glycopeptides are modified after synthesis by enzymes such as glycosyltransferases in solution or in situ on the microarray slide. The constructed glycopeptide microarray is incubated with interaction partners, followed by the detection of the binding event by fluorescence detection or label-free methods, such as mass spectrometry or surface plasmon resonance imaging.
Figure 1. An example of the glycopeptide microarray workflow
Glycopeptide microarray has the unique ability to map antibody-binding determinants and antibody recognition profiles. It provides an opportunity for efficient screening of enzyme substrate specificity or for determination of enzymatic glycosylation sites. Glycopeptide microarray has proven to be a high-throughput and powerful tool for antibody binding epitope analysis in vaccine development as well as for clinical biomarker discovery. It is potentially a valuable tool to investigate antibodies for immunotherapy and immunodiagnostics. Glycopeptide microarray has been employed in the study of enzyme activities and substrate specificities, the characterization of glycan binding proteins, the mapping of antibody epitopes, the detection of autoantibodies and serodiagnosis.
As one of the leading companies in the omics field with over years of experience in omics study, Creative Proteomics provides Glycopeptide Microarray Assay customized to your needs.
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