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Multivalent binding of concanavalin A on variable-density mannoside microarrays.

Daniel J Valles1, Yasir Naeem, Angelica Y Rozenfeld

  • 1The PhD Program in Chemistry, Graduate Center of the City University of New York, 365 5th Ave, New York, NY 10016, USA. Daniel.Valles@asrc.cuny.edu.

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Summary
This summary is machine-generated.

Researchers developed a new method to control glycan density on surfaces, revealing how this density impacts binding with glycan-binding proteins (GBPs) crucial for cell communication and immunity.

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Area of Science:

  • Glycobiology
  • Biochemistry
  • Surface Chemistry

Background:

  • Cell surface glycans and glycan-binding proteins (GBPs) mediate critical biological processes like immunity and cell communication.
  • Multivalent interactions in the dense glycocalyx are key to binding affinity and selectivity, but quantitative analysis is challenging.
  • Existing methods lack the ability to systematically control glycan density for studying these interactions.

Purpose of the Study:

  • To develop a novel method for fabricating glycan arrays with controllable and systematic variations in glycan density.
  • To quantitatively investigate the effect of glycan density on the avidity of glycan-binding proteins.
  • To model the relationship between glycan spacing and binding affinity.

Main Methods:

  • Fabrication of mannoside glycan arrays using thiol-ene click chemistry and a digital micromirror device-based photochemical printer.
  • Controlled variation of mannoside density within array features by dilution with allyl alcohol.
  • Quantitative measurement of concanavalin A (ConA) binding to immobilized mannosides using fluorescence microscopy.

Main Results:

  • A nonlinear decrease in ConA binding was observed with increasing spacer (decreasing mannoside) concentration.
  • A model was developed correlating average mannoside-mannoside spacing to the observed binding drop-off.
  • The study successfully demonstrated a method to tune glycan density and probe GBP interactions.

Conclusions:

  • Controlled modulation of glycan density is critical for understanding multivalent interactions.
  • Advances in microscale photolithography can mimic glycocalyx architecture for studying cell surface information trafficking.
  • This approach provides a new tool for quantitative glycobiology research.