PET Foils Functionalized with Reactive Copolymers as Adaptable Microvolume ELISA Spot Array Platforms for Multiplex...

Area of Science:

• Biomaterials Science
• Diagnostic Assay Development
• Immunotechnology

Background:

• Microvolume ELISA platforms are crucial for high-throughput diagnostics requiring minimal samples.
• Advanced substrate surfaces are essential for efficient biomolecule immobilization and antifouling properties, enhancing assay sensitivity and specificity.

Purpose of the Study:

• To develop and evaluate novel PET-based microvolume ELISA spot arrays for sensitive and specific antibody detection.
• To investigate the efficacy of amine- and DBCO-reactive copolymers for biomolecule immobilization and reduced non-specific binding.

Main Methods:

• Fabrication of PET microvolume ELISA spot arrays coated with MCP-2 and Copoly Azide copolymers.
• Covalent attachment of SARS-CoV-2 nucleoprotein (NP) for antibody immobilization.
• Validation using classical ELISA and Surface Plasmon Resonance (SPR) analysis.
• Clinical evaluation with human serum samples for anti-NP antibody detection.

Main Results:

• Covalent attachment demonstrated superior efficiency and reduced non-specific binding compared to passive adsorption.
• The arrays enabled sensitive detection of IgG and IgA antibodies, including IgG subclasses, in human serum.
• Clinical testing achieved 100.0% sensitivity and 92.9% specificity for anti-NP antibody detection in COVID-19 samples.
• DNA-directed immobilization facilitated multiplex serological measurements.

Conclusions:

• PET-based microvolume ELISA arrays with advanced surface coatings provide a scalable, high-throughput diagnostic solution.
• The platform demonstrates significant potential for sensitive and specific detection of multiple biomarkers, including COVID-19 antibodies.
• Integration into microfluidic devices is feasible, paving the way for advanced diagnostic applications.