Carbohydrate microarrays preparation

In a similar method, Lee and Shin (36) have developed hydrazide-coated glass slides to immobilize a wide range of carbohydrates including mono-, di-, and oligosaccharides in a simple, efficient, and chemoselective fashion. Preliminary protein-binding experiments show that carbohydrate microarrays prepared by this method are suitable for the high-throughput analysis of carbohydrate-protein interactions. 

Figure 11 Carbohydrate microarrays prepared by covalent immobilization of maleimide-derivatized carbohydrates onto a thiol-functionalized substrate surface.

Lee M, Shin I. Eacile preparation of carbohydrate microarrays by site-specific, covalent immobilization of unmodified carbohydrates on hydrazide-coated glass slides. Org. Lett. 2005 7 4269-4272. 

Hitherto, many methods for the preparation of carbohydrate microarrays have been described, such as nitrocellulose coated slides for noncovalent immobilization of microbial polysaccharides [23], and self-assembled monolayers modified by Diels-Alder mediated coupling of cydopentadiene-derivatized oligosaccharides [24], just to name two. Unfortunately, the first method requires large polysaccharides or lipid modified sugars for the noncovalent interaction. The latter method requires the preparation of oligosaccharides bearing the sensitive cyclopentadiene moiety. 

Prepare the carbohydrate microarray immobilized by carbohydrate probes whose binding affinities are determined, according to the procedure described in Subheading 3.3.I.I. 

Prepare the carbohydrate microarray containing GlcNAc according to procedure described in Subheading 3.3.1.1 (see Fig. 5). 

Noncovalent adsorption of native carbohydrate probes on a substrate surface is the simplest way to prepare carbohydrate microarrays. This method relies on the formation of a variety of noncovalent interactions between the surface and the arrayed carbohydrates. In addition to its simplicity and high-throughput characteristics in array construction, these approaches may be favorable in supporting the preservation of the native structure of spotted carbohydrate antigens since there is no need to modify the carbohydrates before microarray application. However, given that the saccharides are noncovalently immobilized on an array substrate, the efficiency of immobilization must be verified for each spotted carbohydrate. 

A surface-modified polystyrene substrate provides another type of polymer surface that can be directly used to prepare carbohydrate microarrays through... 

Overall, these results demonstrated that the binding of lectins with the prepared carbohydrate microarrays are specific and multiple-analytic characterization can be achieved on the aminooxyacetyl-functionalized slide with good selectivity. Furthermore, periodate oxidation of the immobilized oligosaccharides with NaI04 resulted in the loss of lectin binding. These experiments verify that the fabricated carbohydrate microarray is well suited for the selective identification of carbohydrate-binding proteins. 

Zhi ZL, Powell AK, Turnbull JE. Fabrication of carbohydrate microarrays on gold surfaces direct attachment of nonderivatized oligosaccharides to hydra-zide-modified self-assembled monolayers. Anal Chem 2006 78 4786-4793. Lee M, Shin I. Facile preparation of carbohydrate microarrays by site-specific, covalent immobilization of unmodified carbohydrates on hydrazide-coated glass slides. Org Lett 2005 7 4269-4272. 

Wang, H., Y. Zhang, X. Yuan, Y. Chen, and M. Yan. 2011. A universal protocol for photochemical covalent immobilization of intact carbohydrates for the preparation of carbohydrate microarrays. Bioconjug. Chem. 22 26-32. 

Structural and conformational analysis which complements aspects in Section 4 of carbohydrate-protein interaction at the macromolecular level. This is due to an increased number of studies on the preparation of ligands for combinatorial chemistry and the design of carbohydrate microarrays (a recently popular activity coming of age along with genomic and proteomic methods). 

Liu Y, Chai W, Childs RA, Feizi T. Preparation of neoglycol-ipids with ring-closed cores via chemoselective oxime-Ugation for microarray analysis of carbohydrate-protein interactions. Methods Enzymol. 2006 415 326-340. 

General procedures to manufacture, application and management of microarrays have been described in section 14.3. Special concerns to prepare carbohydrate micioar-rays will be dealt with in this subsection. Two methods are used to immobilize glycans on the surface for the fabrication of glycochips (Feizi et al., 2003) noncovalent and covalent immobilization technologies. 

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