Journal: Scientific Reports

Article Title: Integrated Microfluidic Lectin Barcode Platform for High-Performance Focused Glycomic Profiling

doi: 10.1038/srep20297

Figure Lengend Snippet: ( A ) Schematic illustration of the chip design and the scheme of antibody-lectin sandwich assay (inset). The two-layer PDMS chip integrates eight parallel units each consisting of a three-valve pump and an actuatable assay chamber. The assembled chip is sealed onto a glass slide with the assay chambers aligned vertically across an array of lectins patterned on the substrate surface. Glycoproteins are captured by the arrayed lectins and detected by biotinylated antibodies and fluorescently labeled streptavidin. Pneumatic actuation of the assay chamber promotes fluidic mixing for fast affinity binding. ( B ) Digital photo of an assembled chip filled with red food dye in the bottom flow channels and green dye in the pneumatic control channels.

Article Snippet: Biotinylated BSA (Vector Labs) was used as positive control for lectin patterning.

Techniques: Labeling, Binding Assay

Journal: Scientific Reports

Article Title: Integrated Microfluidic Lectin Barcode Platform for High-Performance Focused Glycomic Profiling

doi: 10.1038/srep20297

Figure Lengend Snippet: ( A ) Binding of the glycosylated detection antibody on the arrayed lectins can cause severe non-specific background. To mitigate this effect, the target-bound lectin array is blocked with a non-labeled IgG blocker prior to detection using a biotinylated antibody and fluorescently labeled streptavidin. ( B ) The predominant glycan structures on the standard glycoproteins used: bovine RNase B and human transferrin (hTf). The square brackets indicate the possible high mannose variants on RNase B. ( C , D ) False-color fluorescence images of glyco-profiling of bovine RNase B ( C ) and hTf ( D ) using a 16-lectin barcode array and different blockers-BSA, human plasma IgG (hIgG) and rabbit serum IgG (rIgG). The concentrations of RNase B and hTf are 5 and 50 μg/mL, respectively.

Article Snippet: Biotinylated BSA (Vector Labs) was used as positive control for lectin patterning.

Techniques: Binding Assay, Labeling, Fluorescence