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Aldehyde coupling

Aldehyde coupling provides an alternative approach for immobilizing glycoproteins and other glycoconjugates. The method is particularly suitable for ligands containing sialic acid, since these residues are very easily oxidized to aldehydes using sodium periodate.

Chemistry

Aldehyde coupling immobilizes ligands containing aldehyde groups (either native or introduced by oxidation of cis-diols) by reaction with carbohydrazide on the sensor chip surface. This approach is useful for carbohydrates and glycoproteins.

Prior to injection of the ligand, excess reactive esters are deactivated with ethanolamine. Sodium cyanoborohydride is used to stabilize the linkage of the ligand to the surface.

Aldehyde coupling chemistry

Immobilization procedure overview

The general pattern of steps involved in immobilizing ligand is the same for essentially all covalent immobilization methods:

• Activate the surface by injection of appropriate reagents.
• Inject the ligand solution.
• Inject reagent to deactivate remaining active groups on the surface and remove non-covalently bound ligand.

There may be additional steps depending on the properties of the ligand, the type of sensor surface and the details of the chemistry used.

Lab protocol

Lab procedure Aldehyde coupling of ligand to Biacore sensor chips is available for download on www.cytivalifesciences.com/solutions/protein-research/Interaction-analysis-with-Biacore-surface-plasmon-resonance-SPR/Get-started-with-surface-plasmon-resonance-SPR-interaction-analysis

See also

List of help files

Procedure for oxidizing the ligand