Near real-time biosensor-based detection of 2,4-dinitrophenol.

Area of Science:

• Analytical Chemistry
• Biotechnology
• Immunosensor Technology

Background:

• 2,4-dinitrophenol (DNP) is an explosive and toxic compound requiring sensitive detection methods.
• Existing detection methods may lack the speed and sensitivity required for real-time monitoring.
• Immunoassay techniques offer high specificity for analyte detection.

Purpose of the Study:

• To develop a novel fluorescent biosensor assay for rapid and sensitive detection of 2,4-dinitrophenol (DNP).
• To utilize the KinExA immunoassay instrument for antibody/antigen interaction analysis in solution.
• To establish a versatile platform adaptable for detecting other target analytes.

Main Methods:

• Development of a fluorescent biosensor assay based on antibody/antigen interactions.
• Utilized the KinExA instrument with a capillary flow observation cell and antigen-coated beads (DNP-HSA conjugate on PMMA beads).
• Incubation of samples with anti-DNP monoclonal antibody, followed by binding to antigen-coated beads and detection with a FITC-labeled secondary antibody.

Main Results:

• The developed assay demonstrated near real-time detection of DNP.
• The assay achieved a minimum detection limit of 5 ng/ml for DNP in solution.
• Fluorescence measurements, either from slopes or retained fluorescence, correlated with DNP concentration.

Conclusions:

• A sensitive and rapid fluorescent biosensor assay for DNP detection has been successfully developed.
• The assay leverages the KinExA instrument and antibody/antigen binding principles.
• The platform is adaptable for the detection of various analytes by modifying antigen and antibody pairs.