A Universal Protocol for Photochemical Covalent Immobilization of Intact Carbohydrates for the Preparation of Carbohydrate Microarrays

Abstract

A universal photochemical method has been established for the immobilization of intact carbohydrates and their analogues, and for the fabrication of carbohydrate microarrays. The method features the use of perfluorophenyl azide (PFPA)-modified substrates and the photochemical reaction of surface azido groups with printed carbohydrates. Various aldoses, ketoses, non-reducing sugars such as alditols and their derivatives can be directly arrayed on the PFPA-modified chips. The lectin-recognition ability of arrayed mannose, glucose and their oligoand polysaccharides were confirmed using surface plasmon resonance imaging and laser-induced fluorescence imaging.

Lectin-gated, mesoporous, photofunctionalized glyconanoparticles for glutathione-responsive drug delivery

“…Disulfide formation was supported by the azide group absorption at 2175 cm −1 in the FTIR spectrum, accompanied by the disappearance of the Raman thiol signal (Figure S2). The particles were finally mannosylated under UV irradiation using a previously developed photocoupling method, 18 where the PFPA azide moiety is converted into a nitrene that can undergo insertion into carbohydrate to give D-mannose-conjugated nanoparticles ( FMSN-Man , Scheme 1). The carbohydrate content in FMSN-Man was estimated by an anthrone-based colorimetry assay, 17b,19 indicating an approximate amount of 12 μg of D-mannose per mg of particles (Figure S3), and a coupling yield of 48%.…”

Microarray-Based Technology for Glycomics Analysis

“…Recently, several microarray platforms with nonfluorescence detection techniques including mass spectrometry (MS) [25,39,78,79], surface plasmon resonance (SPR) [52,53,[80][81][82][83], scanning ellipsometry (imaging) [84], and resonant light scattering (RLS) of nanoparticle [85,86], have developed for glycomic analysis. For instance, Cheng and coworkers reported the fabrication of a carbohydrate microarray with surface plasmon resonance imaging (SPRi) detection system for studying interactions of carbohydrates with lectins (see Fig.…”

“…On the other hand, covalent binding of carbohydrates with substrates offers high stability and is demonstrated to be quite robust [39][40][41][42][43][44][45][46][47][48][49][50][51][52][53]. Therefore, the most general method for construction of these microarrays involves site-specific and covalent immobilization of carbohydrates to properly derivatized surfaces.…”

“…This immobilization procedure is generally applicable to reductively aminated glycans with different established fluorescent labels and allows the spatial arrangement of oligosaccharides. Chen and coworkers reported a universal photochemical method for the immobilization of intact carbohydrates and their analogues on perfluorophenyl azide (PFPA)-modified substrates by the photochemical reaction of surface azido groups with printed carbohydrates [53]. The immobilization takes the advantage of solid-phase reaction rather than solution reaction.…”

Analysis of the surface density and reactivity of perfluorophenylazide and the impact on ligand immobilization

“…The versatility of the PFPA chemistry makes it an attractive choice for surface modification upon thermal activation or light irradiation of the azide functionality. [1][2][3][4][5][6][7][8][9][10][11][12] The PFPA immobilization method is simple, reproducible, and has been proven to be efficient to immobilize synthetic polymers, 1 graphene, 2,3 and carbohydrates [4][5][6][7] onto variety of surfaces. 8 Moreover, PFPA allows controlling the surface density of the immobilized molecules through adjustment of the PFPA solution concentration.…”