Production and analytical characterization of neopterin immunoreagents for biosensor developments

Area of Science:

• Immunology
• Biochemistry
• Biomarker Research

Background:

• Neopterin is a key biomarker for cellular immunity, indicating various pathological conditions like infections and cancers.
• Accurate and rapid detection of neopterin is crucial for timely diagnosis and disease management.
• Existing detection methods may lack the sensitivity or speed required for multi-biomarker applications.

Purpose of the Study:

• To synthesize novel neopterin derivatives for antibody production.
• To generate and characterize high-affinity monoclonal and polyclonal antibodies against neopterin.
• To develop and validate enzyme-linked immunosorbent assay (ELISA) methods for neopterin detection.

Main Methods:

• Synthesis of a novel hapten derivative of neopterin and conjugation to carrier proteins.
• Immunization of mice and rabbits to produce monoclonal and polyclonal antibodies.
• Characterization of antibody performance using direct and indirect ELISA, including limit of detection (LoD) determination.
• Validation of ELISA methods with clinical plasma samples from various patient groups.

Main Results:

• Successfully generated monoclonal and polyclonal antibodies against neopterin with high specificity and affinity.
• Achieved low limits of detection (LoD) for neopterin: 0.18 µg/L (monoclonal, direct ELISA) and 0.05 µg/L (polyclonal, direct ELISA).
• Demonstrated reliable neopterin quantification in human plasma samples (3.2–103 µg/L) using the developed ELISA methods.
• Observed satisfactory correlation between the developed ELISA methods and a commercial kit.

Conclusions:

• The generated neopterin antibodies are suitable for sensitive and specific detection.
• These antibodies can be integrated into rapid, multi-biomarker diagnostic platforms and biosensor systems.
• The developed ELISA methods provide valuable tools for clinical diagnostics and research involving neopterin.