N-Glycan Array 100

The 100 N-Glycan Array is used as a general test to help researchers determine binding characteristics of antibodies, proteins, bacteria, cell cultures, or other potential biological samples to an array of 100 fundamental N-glycans. Our microarray slides are especially coated to be capable of immobilizing natural N-glycans with close-ring structure at their reducing end (GlcNAc). N-linked oligosaccharides in particular play a major role in intercellular interactions and immune cell functions, making them often preferred glycan candidates for the study of carbohydrate-binding-vaccines such as HIV Broadly Neutralizing Antibodies (bNAbs).  The fundamental N-glycan structures provided in this array can provide understanding of basal binding determinants for antibodies or other proteins of interest.

N-Glycan Features

How Pre-complexing Assay Works

Certain broadly neutralizing antibodies need a pre-complexing assay to enhance their relatively weak binding to N-glycans. Below is an illustration for the pre-complexing binding assays:

General Lectin Binding Characteristics to N-Glycans

Lectins are proteins with binding pockets that recognize specific aspects of glycans. This image shows the basic binding characteristics of common lectins.  Due to the lectins’ natural preference for multivalent binding, binding affinity is often affected by the properties of the glycan as a whole including structure, components, and how accessible the glycan is to the lectin. These N-glycans displayed are among the most strongly binding N-glycans to each respective lectin.

Typical Binding Assay Result from the 100 N-Glycan Array

Example 1:  A subarray assayed with glycan-binding protein biotinylated ConA lectin, followed by streptavidin-Cy3 conjugate. Array was scanned with a GenePix scanner at 475 PMT and 100% laser power at 532nm wavelength. There is no non-specific binding for the negative control spots. A positive control and the marker show binding as expected, as well as mannose-containing N-glycans.

Example 2:  A subarray assayed with glycan-binding protein biotinylated AAL lectin, followed by streptavidin-Cy3 conjugate. Array was scanned with a GenePix scanner at 475 PMT and 100% laser power at 532nm wavelength. There is no non-specific binding for the negative control spots.  A positive control and the marker show binding as expected, as well as fucosylated N-glycans.