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Development of lectin arrays for mapping the glycosylation changes in proteins and cells

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KOMÁREK Jan KORSÁK Marek FALTINEK Lukáš WIMMEROVÁ Michaela

Rok publikování 2022
Druh Konferenční abstrakty
Fakulta / Pracoviště MU

Přírodovědecká fakulta

Citace
Popis Saccharides are essential for life, and in addition to their traditional roles as structural and energy storage components, they play crucial roles in many recognition events, signaling, and communication processes. The glycosylation of cells reflects the biological species, tissue, and physiological state of the organism. Importantly, changes in glycosylation patterns were observed in various tumors, and the aberrant glycosylation is often associated with malignancy potential, tumor immune surveillance, and patients´ prognosis. Lectin arrays are widely used tools for analyzing glycosylation in proteins and cells. They utilize a panel of lectins (saccharide-binding proteins with distinct sugar specificities) immobilized onto a solid surface. Lectin arrays allow for rapid and high-throughput glycomic analysis with minimal sample consumption. They can be used for glycan profiling of various samples, including purified glycoproteins, cell/tissue lysates, membrane vesicles, complex biological samples, or entire cells. This talk will present the development of customized lectin arrays in two different formats. The classical array in the microscope slide format was prepared by non-contact piezo dispensing of lectins onto a functionalized glass slide and their immobilization using various surface chemistries (immobilization on epoxy groups, NHS/EDC coupling). In the slide format, binding of glycosylated samples was detected by fluorescence measurement. To allow label-free detection of attached glyco-samples, lectin microarray prepared on bio-layer interferometry (BLI) sensor tips is currently being optimized. The fabricated lectin arrays will be used to map the glycosylation changes at different stages of cell differentiation and under different physiopathological conditions.
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