See Image/sAfter biotinylation of HRP we can determine what kind of glycans are conjugated to HRP by using the RayBiotech lectin array. Lectins, GNA, HHA, NPA showed strong signals after incubation with 0.33 ug/mL Biotin-HRP followed detection by streptavidin-fluorescence-dye (Figure A, B and C). The fluorescence signals from GNA, HHA and NPA can be blocked in a concentration depend manner by HRP itself (Figure A and B), which means that these fluorescence signals were generated based on the binding between HRP and the three lectins. As we know, GNA, HHA and NPA lectins specifically bind to mannose which indicates that HRP contains mannose. By adding increasing amount of mannose, the signal from GNA, HHA and NPA can be reduced (Figure A and C). The reduction signals from increasing concentrations of mannose confirms that HRP protein contains mannose in its glycocalyx. Lectin VVA binds to the streptavidin-fluorescence dye.

See Image/sUsing the lectin array, we can discover the different glycoprotein profiles of the serum samples or cell lysates from patient cohorts versus a control group. Below images showed the profiles of the glycans from serum samples detected by Biotin-anti-human IgG and Fluorescence dye-streptavidin.

Quantitative analysis of lectin-glycoprotein interactions. Example: a concentration series of glycoproteins detected with the lectin array could reveal concentration dependent effects of lectin-glycan binding.Determine the profile of bacterial cell-surface glycans. Example: Cell lysate from bacteria can be Biotinylated and hybridized to the lectin array. Analysis of the binding pattern and correlation with the known carbohydrate-binding specificities of the lectins can determine the glycans on the cell membrane.