Affinity-Based Copolymer Coating for Oriented Protein Immobilization in Biosensor Development

Area of Science:

• Materials Science
• Biotechnology
• Analytical Chemistry

Background:

• Protein immobilization is crucial for biosensor stability and functionality.
• Existing methods often lack control over protein orientation and can reduce activity.
• There is a need for advanced immobilization strategies for next-generation biosensors.

Purpose of the Study:

• To develop and validate a novel affinity-based terpolymer coating for enhanced protein immobilization.
• To investigate the binding capacity, stability, and reversibility of the new coating for histidine-tagged proteins.
• To assess the impact of the coating on enzyme activity after immobilization.

Main Methods:

• Synthesis of a terpolymer incorporating nitrilotriacetic acid (NTA) ligands.
• Coating magnetic microbeads with the novel terpolymer.
• Testing protein binding (native and His-tagged) and enzyme activity compared to traditional amine-reactive coatings.

Main Results:

• The NTA-functionalized terpolymer demonstrated superior binding capacity for histidine-tagged proteins via metal-chelating interactions.
• The immobilization exhibited enhanced stability and reversibility compared to amine-reactive methods.
• Immobilized enzymes retained their activity, indicating minimal denaturation.

Conclusions:

• The novel affinity-based terpolymer coating provides an effective, versatile, and scalable solution for protein immobilization in biosensors.
• This approach facilitates oriented binding and preserves biomolecular function, crucial for sensitive and reliable diagnostic tools.
• The technology holds significant potential for advancing biosensor platforms and analytical technologies.