Z Biotech’s 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 fundamental N-glycans. 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. The fundamental N-glycan structures provided in this array can provide understanding of basal binding determinants for antibodies or other proteins of interest. This manual is provided as a comprehensive guide to help the researcher acquire clear results from the assay. Please read through carefully before starting your experiment.
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 N-Glycan Array
The N-glycan Array was assayed with biotinylated ConA lectin, followed by the streptavidin-Cy3 conjugate. The array was scanned with a microarray scanner 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.