DNA-directed immobilisation of glycomimetics for glycoarrays application: Comparison with covalent immobilisation, and development of an on-chip IC50 measurement assay

Zhang, Jing; Pourceau, Gwladys; Meyer, Albert; Vidal, Sébastien; Praly, Jean‐Pierre; Souteyrand, Éliane; Vasseur, Jean‐Jacques; Morvan, François; Chevolot, Yann

doi:10.1016/j.bios.2009.01.007

Cited by 44 publications

(49 citation statements)

References 53 publications

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“…[37,40] In particular, IC 50 and K d assaysb ased on DDI microarraysf or quantitative analysiso fl ectin-glycocluster interactions were developed. [38,43] However, the impact of glycocluster surface densities on measurements of IC 50 and K d values has to the best of our knowledge not previously been reportedi nt he literature. This is of crucial importance,e specially in the case of lectin-glycocluster interactions, due to the cluster effect.…”

Section: Resultsmentioning

confidence: 91%

Effects of the Surface Densities of Glycoclusters on the Determination of Their IC₅₀ and K_d Value Determination by Using a Microarray

et al. 2015

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Pseudomonas aeruginosa (PA) is an opportunistic bacterium involved in 10-30% of nosocomial diseases. It causes severe lung injury to cystic fibrosis patients, often leading to patient death. PA strains are multidrug resistant, thus making the design of new therapeutics a challenge for public health. One promising therapeutic option is to design glycoclusters that target the virulence factor of PA. LecA is a galactose-specific lectin that might be involved in adhesion and biofilm formation by PA. The DNA-directed immobilization (DDI) microarray is a powerful tool for screening and understanding of structure-activity relationships between glycoclusters and lectins. High-throughput and multiplexed analysis of lectin-glycocluster interactions on a DDI microarray allows measurement of IC50 and dissociation constant (Kd ) values with minute amounts of material. In order to study the robustness of the DDI microarray in determination of IC50 and Kd values, the impact of glycocluster surface density was investigated. The data obtained show that measured IC50 values were influenced by glycocluster surface density: as the density of glycoclusters increases, the measured IC50 values increase too. In contrast, the measured Kd values were not affected by glycocluster surface density, provided that the experimental conditions allow interaction between glycocluster and lectin at single-molecule level (no surface cluster effect).

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Section: Resultsmentioning

confidence: 91%

Effects of the Surface Densities of Glycoclusters on the Determination of Their IC₅₀ and K_d Value Determination by Using a Microarray

et al. 2015

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“…The IC 50 values for the monogalactose 24 a, [13] the trigalactose 24 b, [13] the tetragalactose 22, and the octagalactose 23 glycoconjugates are displayed in Table 1.…”

Section: Synthesis and Characterization Of Glycoclusteroligonucleotidmentioning

confidence: 99%

“…This good result is very likely due to the postulated [25] complexation of a copper(I)-triazolide intermediate with the sugar alkyne, which is therefore placed in proximity to the unreacted azido groups and can readily undergo an intramolecular cycloaddition. The regioisomeric assignment of the 1,4-disubstituted 1,2,3-triazole rings was established by 13 C NMR spectroscopy as described in our earlier works.…”

Section: Wwwchembiochemorgmentioning

confidence: 99%

“…[13] We determined IC 50 values as the concentration of lactose required for the removal of 50 % of RCA 120 bound to the immobilized glycoconjugates. The glycoconjugates 22 or 23 were immobilized by hybridization at the bottoms of the microwells and further incubated with mixtures of Cy5-labeled RCA 120 (2 mm) containing increasing concentrations of lactose (0.05 mm to 30 mm).…”

Section: Synthesis and Characterization Of Glycoclusteroligonucleotidmentioning

confidence: 99%

“…[10,11] The evaluation of the affinity of each glycomimic was performed on a DNA-based glycoarray platform as previously described [12] by direct fluorescence scanning and by determination of its IC 50 value. [13] Interactions between proteins and carbohydrates are involved in a large number of crucial biological events. Many efforts have been devoted to the design and synthesis of unnatural saccharides displaying high affinities towards targeted lectins.…”

Section: Introductionmentioning

confidence: 99%

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Design of Triazole‐Tethered Glycoclusters Exhibiting Three Different Spatial Arrangements and Comparative Study of their Affinities towards PA‐IL and RCA 120 by Using a DNA‐Based Glycoarray

et al. 2009

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Sugar-coated chips: Glycoside clusters are valuable tools for carbohydrate-lectin recognition studies. However, the spatial arrangement of the sugar residues is a key issue in the design of high-affinity glycoclusters. Here the affinities of linear and antenna- and calixarene-based galactoside clusters towards two lectins derived from Pseudomonas aeruginosa and Ricinus communis were compared by means of glycoarrays.Interactions between proteins and carbohydrates are involved in a large number of crucial biological events. Many efforts have been devoted to the design and synthesis of unnatural saccharides displaying high affinities towards targeted lectins. Among others, glycoside clusters have proven to be valuable tools for these recognition studies. However, the spatial arrangements of the sugar residues are a key issue in the design of high-affinity glycoclusters. Here, the affinities of linear and antenna- and calixarene-based galactoside clusters against two lectins, derived from Pseudomonas aeruginosa and Ricinus communis, have been compared by means of glycoarrays.

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Carbohydrate‐Oligonucleotide Conjugates

Pourceau

Meyer

Vasseur

et al. 2009

CP Nucleic Acid Chemistry

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This unit describes a strategy for attaching two mannose and two galactose residues to an oligonucleotide. This conjugation can be performed at the 5'-end of the oligonucleotide sequence, using modified phosphoramidites. First, the oligonucleotide scaffold is synthesized on solid support using a DNA synthesizer, with commercially available and modified phosphoramidites. After the first "click" reaction with a galactosylated azide derivative on solid support, the bromine atoms are replaced with azides and a second click reaction is performed with propargylated mannose either on solid support or in solution. Additionally, using a monoalkynated solid support, the conjugation with carbohydrate residues can be performed at the 3'-end of the oligonucleotide according to a similar protocol. Curr. Protoc. Nucleic Acid Chem. 39:4.38.1-4.38.25.

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DNA-directed immobilisation of glycomimetics for glycoarrays application: Comparison with covalent immobilisation, and development of an on-chip IC50 measurement assay

Cited by 44 publications

References 53 publications

Effects of the Surface Densities of Glycoclusters on the Determination of Their IC₅₀ and K_d Value Determination by Using a Microarray

Effects of the Surface Densities of Glycoclusters on the Determination of Their IC₅₀ and K_d Value Determination by Using a Microarray

Design of Triazole‐Tethered Glycoclusters Exhibiting Three Different Spatial Arrangements and Comparative Study of their Affinities towards PA‐IL and RCA 120 by Using a DNA‐Based Glycoarray

Carbohydrate‐Oligonucleotide Conjugates

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DNA-directed immobilisation of glycomimetics for glycoarrays application: Comparison with covalent immobilisation, and development of an on-chip IC50 measurement assay

Cited by 44 publications

References 53 publications

Effects of the Surface Densities of Glycoclusters on the Determination of Their IC50 and Kd Value Determination by Using a Microarray

Effects of the Surface Densities of Glycoclusters on the Determination of Their IC50 and Kd Value Determination by Using a Microarray

Design of Triazole‐Tethered Glycoclusters Exhibiting Three Different Spatial Arrangements and Comparative Study of their Affinities towards PA‐IL and RCA 120 by Using a DNA‐Based Glycoarray

Carbohydrate‐Oligonucleotide Conjugates

Contact Info

Product

Resources

About

Effects of the Surface Densities of Glycoclusters on the Determination of Their IC₅₀ and K_d Value Determination by Using a Microarray

Effects of the Surface Densities of Glycoclusters on the Determination of Their IC₅₀ and K_d Value Determination by Using a Microarray