On-Chip Neo-Glycopeptide Synthesis for Multivalent Glycan Presentation

Area of Science:

• Carbohydrate chemistry
• Biophysical chemistry
• Glycobiology

Background:

• Glycan-protein interactions are crucial in biology but often weak individually.
• Multivalent interactions, using multiple binding sites, enhance avidity and specificity.
• Existing array technologies often lack precise control over multivalent display.

Purpose of the Study:

• To develop a method for synthesizing well-defined multivalent glycan scaffolds on microarrays.
• To investigate the impact of glycan density and spacing on lectin binding.
• To enable rapid screening and optimization of multivalent glycan-binding agents.

Main Methods:

• Utilized laser-based array synthesis for on-surface peptide synthesis.
• Employed click chemistry to create neo-glycopeptides directly on functionalized glass slides.
• Developed microarrays with tunable carbohydrate density and spatial distribution.
• Probed lectin binding (concanavalin A, langerin) using the synthesized multivalent scaffolds.

Main Results:

• Successfully generated neo-glycopeptides in a microarray format with controlled glycan presentation.
• Demonstrated that lectin binding is dependent on glycan spacing, density, and the specific lectin.
• Enabled measurement of surface dissociation constants for glycan-lectin interactions.
• Showcased the ability to rapidly generate and screen diverse multivalent scaffolds.

Conclusions:

• The developed laser-based synthesis and click chemistry approach allows for precise control over multivalent glycan display.
• This method significantly advances the study of glycan-protein interactions by enabling tunable scaffold generation.
• The platform facilitates rapid screening and optimization of multivalent structures for glycan binding applications.