Label-Free Discovery Array Platform for the Characterization of Glycan Binding Proteins and Glycoproteins

Gray, Christopher J, Sanchez-Ruiz, Antonio, Sardzikova, Ivana, Ahmed, Yassir A, Miller, Rebecca L, Reyes Martinez, Juana E, Pallister, Edward, Huang, Kun, Both, Peter, Hartmann, Mirja
et al (show 6 more authors) (2017) Label-Free Discovery Array Platform for the Characterization of Glycan Binding Proteins and Glycoproteins. ANALYTICAL CHEMISTRY, 89 (8). pp. 4444-4451.

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The identification of carbohydrate-protein interactions is central to our understanding of the roles of cell-surface carbohydrates (the glycocalyx), fundamental for cell-recognition events. Therefore, there is a need for fast high-throughput biochemical tools to capture the complexity of these biological interactions. Here, we describe a rapid method for qualitative label-free detection of carbohydrate-protein interactions on arrays of simple synthetic glycans, more complex natural glycosaminoglycans (GAG), and lectins/carbohydrate binding proteins using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. The platform can unequivocally identify proteins that are captured from either purified or complex sample mixtures, including biofluids. Identification of proteins bound to the functionalized array is achieved by analyzing either the intact protein mass or, after on-chip proteolytic digestion, the peptide mass fingerprint and/or tandem mass spectrometry of selected peptides, which can yield highly diagnostic sequence information. The platform described here should be a valuable addition to the limited analytical toolbox that is currently available for glycomics.

Item Type: Article
Uncontrolled Keywords: Milk, Human, Humans, Trypsin, Glycoproteins, Glycosaminoglycans, Peptides, Lectins, Microarray Analysis, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Protein Binding
Depositing User: Symplectic Admin
Date Deposited: 15 Mar 2017 14:25
Last Modified: 19 Jan 2023 07:12
DOI: 10.1021/acs.analchem.6b04122
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