Evaluation of hetero-multivalent lectin binding using a turbidity-based emulsion agglutination assay.

Lectin hetero-multivalency, binding to two or more different types of ligands, has been demonstrated to play a role in case of both LecA (a Pseudomonas aeruginosa adhesin) and Cholera Toxin subunit B (a Vibrio cholerae toxin). In order to screen the ligand candidates that involve in hetero-multivalent binding from large molecular libraries, we present a turbidity-based emulsion agglutination (TEA) assay that can be conducted in a high-throughput format using the standard laboratory instruments and reagents. The benefit of this assay is that it relies on the use of emulsions that can be formed using ultrasonication, minimizing the bottleneck of substrate surface functionalization.

Hetero-Multivalency of Pseudomonas aeruginosa Lectin LecA Binding to Model Membranes.

A single glycan-lectin interaction is often weak and semi-specific. Multiple binding domains in a single lectin can bind with multiple glycan molecules simultaneously, making it difficult for the classic "lock-and-key" model to explain these interactions. We demonstrated that hetero-multivalency, a homo-oligomeric protein simultaneously binding to at least two types of ligands, influences LecA (a Pseudomonas aeruginosa adhesin)-glycolipid recognition.

Hetero-multivalent binding of cholera toxin subunit B with glycolipid mixtures.

GM has generally been considered as the major receptor that binds to cholera toxin subunit B (CTB) due to its low dissociation constant. However, using a unique nanocube sensor technology, we have shown that CTB can also bind to other glycolipid receptors, fucosyl-GM and GDb. Additionally, we have demonstrated that GM can contribute to CTB binding if present in a glycolipid mixture with a strongly binding receptor (GM/fucosyl-GM/GDb).

Quantitative surface-enhanced Raman spectroscopy for kinetic analysis of aldol condensation using Ag-Au core-shell nanocubes.

Surface-enhanced Raman spectroscopy (SERS) is a powerful tool with high potential for multiplexed detection of dilute analytes. However, quantitative SERS of kinetic assays can be difficult due to the variation in enhancement factors caused by changing reaction conditions. We report a method for quantitative SERS kinetic analysis using colloidal Ag-Au core-shell nanocubes (Ag@AuNCs) as the SERS substrate.

Binding Cooperativity Matters: A GM1-Like Ganglioside-Cholera Toxin B Subunit Binding Study Using a Nanocube-Based Lipid Bilayer Array.

Protein-glycan recognition is often mediated by multivalent binding. These multivalent bindings can be further complicated by cooperative interactions between glycans and individual glycan binding subunits. Here we have demonstrated a nanocube-based lipid bilayer array capable of quantitatively elucidating binding dissociation constants, maximum binding capacity, and binding cooperativity in a high-throughput format.