High-throughput SPR biosensor.

Area of Science:

• Biochemistry
• Analytical Chemistry
• Glycomics

Background:

• Surface plasmon resonance (SPR) imaging is a label-free, real-time, high-throughput method for studying molecular interactions in an array format.
• SPR imaging offers potential for screening small molecule arrays in reverse chemical genomics.

Purpose of the Study:

• To demonstrate the fabrication of sugar arrays using photoaffinity crosslinking.
• To investigate sugar-lectin interactions using SPR imaging.
• To establish a protocol for array fabrication without chemical modification of sugar chains.

Main Methods:

• Fabrication of small molecule arrays via photoaffinity crosslinking.
• Utilizing biotinylated sugars for immobilization.
• Employing SPR imaging for real-time interaction analysis.
• Investigating lectin binding to immobilized sugars.

Main Results:

• Demonstrated a protocol for fabricating sugar arrays without chemical modification of sugar chains.
• Successfully studied sugar-lectin interactions using SPR imaging.
• Investigated signal ratios between lectin and antistreptavidin antibody binding for biotinylated sugars.
• Achieved signal normalization despite unclear immobilized sugar densities.

Conclusions:

• SPR imaging is effective for label-free, real-time analysis of sugar-lectin interactions in an array format.
• The demonstrated protocol allows for efficient sugar array fabrication and analysis.
• This method is a promising first screening step for reverse chemical genomics applications.