Piezo dispensed microarray of multivalent chelating thiols for dissecting complex protein-protein interactions

Area of Science:

• Biotechnology
• Surface Chemistry
• Molecular Interactions

Background:

• Understanding multiprotein complex formation is crucial for deciphering cellular processes.
• Existing biochip technologies face limitations in precisely controlling protein immobilization and analyzing complex interactions.

Purpose of the Study:

• To develop a novel biochip for dissecting multiprotein complex formation.
• To characterize the fabrication and functional properties of the biochip for studying protein-protein interactions.

Main Methods:

• Fabrication of a metal chelator array using piezo dispensing of bis-nitrilotriacetic acid (bis-NTA) thiol on gold thin films.
• Characterization of dispensed layers using imaging ellipsometry.
• Demonstration of functional properties via imaging surface plasmon resonance to study type I interferon receptor subunit interactions.

Main Results:

• Dispensed layers showed increased thickness with higher bis-NTA molar fractions compared to solution self-assembly.
• Ligand dissociation kinetics were found to depend on receptor surface concentration, indicating ternary complex formation.
• The biochip successfully enabled multiplexed monitoring of binding phenomena at various receptor densities.

Conclusions:

• The novel biochip fabrication method allows for precise control over protein immobilization density.
• The biochip is effective for dissecting complex protein-protein interactions and analyzing binding kinetics.
• This technology provides a powerful tool for multiplexed analysis of molecular interactions in biological systems.