The interaction of carbohydrates with a variety of biological targets, including antibodies, proteins, viruses and cells are of utmost importance in many aspects of biology. Glycan microarrays are increasingly used to determine the binding specificity of glycan-binding proteins. In this study, a novel slide is reported for the fabrication of glycan arrays that combines the higher sensitivity of a layered Si-SiO<inf>2</inf> with a novel approach to form a polymeric coating easily modifiable by subsequent click reaction. The alkyne-containing copolymer, adsorbed from an aqueous solution, produces a coating by a single step procedure and serves as a soft, tridimensional support for the oriented immobilization of carbohydrates via azide/alkyne Cu(I) catalyzed "click" reaction. The equilibrium and kinetics parameters of the interaction of Concanavalin A with eight synthetic glycans were determined using fluorescence microarray and Reflective Phantom Interface (RPI), a recently proposed optical label-free detection approach. The enhancement of fluorescence provided by the Si-SiO<inf>2</inf> slides enabled to extend the limit of detection at lower surface densities of lectins, in turn enabling the study of the interaction for a wide range of glycans surface density. Equilibrium dissociation constants of a few nM were extracted for multivalent glycan-lectin binding, mimicking the conditions of biological membranes, whereas hundreds of nM were observed at the lower glycan surface densities.

New "clickable" polymeric coating for glycan microarrays / C. Zilio, A. Bernardi, A. Palmioli, M. Salina, G. Tagliabue, M. Buscaglia, R. Consonni, M. Chiari. - In: SENSORS AND ACTUATORS. B, CHEMICAL. - ISSN 0925-4005. - 215:(2015 Aug), pp. 412-420. [10.1016/j.snb.2015.03.079]

New "clickable" polymeric coating for glycan microarrays

C. Zilio
Primo
;
A. Bernardi
Secondo
;
A. Palmioli;M. Salina;G. Tagliabue;M. Buscaglia;
2015

Abstract

The interaction of carbohydrates with a variety of biological targets, including antibodies, proteins, viruses and cells are of utmost importance in many aspects of biology. Glycan microarrays are increasingly used to determine the binding specificity of glycan-binding proteins. In this study, a novel slide is reported for the fabrication of glycan arrays that combines the higher sensitivity of a layered Si-SiO2 with a novel approach to form a polymeric coating easily modifiable by subsequent click reaction. The alkyne-containing copolymer, adsorbed from an aqueous solution, produces a coating by a single step procedure and serves as a soft, tridimensional support for the oriented immobilization of carbohydrates via azide/alkyne Cu(I) catalyzed "click" reaction. The equilibrium and kinetics parameters of the interaction of Concanavalin A with eight synthetic glycans were determined using fluorescence microarray and Reflective Phantom Interface (RPI), a recently proposed optical label-free detection approach. The enhancement of fluorescence provided by the Si-SiO2 slides enabled to extend the limit of detection at lower surface densities of lectins, in turn enabling the study of the interaction for a wide range of glycans surface density. Equilibrium dissociation constants of a few nM were extracted for multivalent glycan-lectin binding, mimicking the conditions of biological membranes, whereas hundreds of nM were observed at the lower glycan surface densities.
Click-chemistry; Glycan; Microarrays; Surface coating
Settore CHIM/06 - Chimica Organica
Settore FIS/07 - Fisica Applicata(Beni Culturali, Ambientali, Biol.e Medicin)
ago-2015
Article (author)
File in questo prodotto:
File Dimensione Formato  
SNB-Submission.pdf

accesso aperto

Descrizione: articolo + figure
Tipologia: Pre-print (manoscritto inviato all'editore)
Dimensione 846.04 kB
Formato Adobe PDF
846.04 kB Adobe PDF Visualizza/Apri
1-s2.0-S0925400515004128-main.pdf

accesso riservato

Tipologia: Publisher's version/PDF
Dimensione 1.1 MB
Formato Adobe PDF
1.1 MB Adobe PDF   Visualizza/Apri   Richiedi una copia
Pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/280039
Citazioni
  • ???jsp.display-item.citation.pmc??? ND
  • Scopus 27
  • ???jsp.display-item.citation.isi??? 24
social impact