Arrayed profiling of multiple glycans on whole living cell surfaces

Area of Science:

• Biochemistry
• Molecular Biology
• Analytical Chemistry

Background:

• Cell surface glycans play crucial roles in biological processes.
• Accurate profiling and quantification of glycans are essential for understanding cellular functions.
• Existing methods for glycan analysis often require cell pretreatment or labeling.

Purpose of the Study:

• To develop an array-based method for profiling and quantifying multiple glycans on whole living cell surfaces.
• To combine DNA encoding technology with DNA microarray for glycan analysis.
• To establish a high-throughput, label-free method for glycomic studies.

Main Methods:

• Development of specific barcode-lectin probes containing endonuclease cutting sites.
• Incubation of living cells with barcode-lectin probes for glycan binding.
• Endonuclease cleavage to release barcodes, followed by DNA microarray hybridization.
• Signal amplification using hybridization chain reaction (HCR) with Cy5-labeled hairpins.

Main Results:

• Successful profiling and quantification of four types of cell surface glycans (mannose, N-acetylgalactosamine, N-acetylglucosamine, N-acetylneuraminic acid).
• Quantification of specific glycan amounts per BGC cell (e.g., 2.1 × 10^8 N-acetylglucosamine moieties/cell).
• Demonstration of whole cell surface accessibility, distinguishing capability, and fast kinetics.

Conclusions:

• The developed array-based method offers a powerful tool for glycan profiling and quantification on living cells.
• The method is high-throughput, miniaturizable, and does not require cell pretreatment or labeling.
• This technique facilitates the elucidation of complex glycan-related biological processes.