Tailored Multivalent Neo-Glycoproteins: Synthesis, Evaluation, and Application of a Library of Galectin-3-Binding Glycan Ligands

0482841 - MBÚ 2018 RIV US eng J - Článek v odborném periodiku
Laaf, D. - Bojarová, Pavla - Pelantová, Helena - Křen, Vladimír - Elling, L.
Tailored Multivalent Neo-Glycoproteins: Synthesis, Evaluation, and Application of a Library of Galectin-3-Binding Glycan Ligands.
Bioconjugate Chemistry. Roč. 28, č. 11 (2017), s. 2832-2840. ISSN 1043-1802. E-ISSN 1520-4812
Grant CEP: GA ČR GC15-02578J; GA MŠMT(CZ) LTC17005
Institucionální podpora: RVO:61388971
Klíčová slova: HOMOTYPIC CELL-AGGREGATION * ONE-POT SYNTHESIS * GALACTOSE-OXIDASE
Obor OECD: Biochemistry and molecular biology
Impakt faktor: 4.485, rok: 2017

Galectin-3 (Gal-3), a member of the beta-galactoside-binding lectin family, is a tumor biomarker and involved in tumor angiogenesis and metastasis. Gal-3 is therefore considered as a promising target for early cancer diagnosis and anticancer therapy. We here present the synthesis of a library of tailored multivalent neo-glycoproteins and evaluate their Gal-3 binding properties. By the combinatorial use of glycosyltransferases and chemo-enzymatic reactions, we first synthesized a set of N-acetyllactosamine (Gal beta,4GlcNAc, LacNAc type 2)-based oligosaccharides featuring five different terminating glycosylation epitopes, respectively. Neo-glycosylation of bovine serum albumin (BSA) was accomplished by dialkyl squarate coupling to lysine residues resulting in a library of defined multivalent neo-glycoproteins. Solid-phase binding assays with immobilized neo-glycoproteins revealed distinct affinity and specificity of the multivalent glycan epitopes for Gal-3 binding. In particular, neo-glycoproteins decorated with N',N '-diacetyllactosamine (GalNAc beta,4GlcNAc, LacdiNAc) epitopes showed high selectivity and were demonstrated to capture Gal-3 from human serum with high affinity. Furthermore, neo-glycoproteins with terminal biotinylated LacNAc glycan motif could be utilized as Gal-3 detection agents in a sandwich enzyme-linked immunosorbent assay format. We conclude that, in contrast to antibody-based capture steps, the presented neoglycoproteins are highly useful to detect functionally intact Gal-3 with high selectivity and avidity. We further gain novel insights into the binding affinity of Gal-3 using tailored multivalent neo-glycoproteins, which have the potential for an application in the context of cancer-related biomedical research.
Trvalý link: http://hdl.handle.net/11104/0278236