Development of a reversible bienzymatic system in immunoenzymatic technique

Area of Science:

• Biotechnology
• Analytical Chemistry
• Immunology

Background:

• Enzyme-linked immunoassays (ELISA) are widely used for detecting analytes.
• Existing ELISA methods can be time-consuming and require manual processing.
• There is a need for automated, sensitive, and rapid immunoassay techniques.

Purpose of the Study:

• To develop and validate a novel, automated enzyme-linked immunoassay (ELISA) for the determination of haptens and antigens.
• To integrate specific captors, enzyme labeling, and a real-time detection system for enhanced assay performance.
• To demonstrate the applicability of the developed method in biological and industrial fluid analysis.

Main Methods:

• Utilized specific captors solubilized with a ligand on proteic membranes.
• Employed a two-enzyme system: one for immunocomplex linking and beta-d-glucose oxidase for antigen labeling.
• Integrated a pO(2) sensor for real-time measurement of oxygen consumption and an automated microcomputer system for signal processing.

Main Results:

• The developed automated ELISA system demonstrated the ability to determine various haptens and antigens.
• Real-time measurement of oxygen consumption by beta-d-glucose oxidase provided a quantifiable signal.
• Signal correlation with urea solution injection confirmed the reliability of the automated system.

Conclusions:

• The proposed automated ELISA technique offers a sensitive and efficient method for analyte determination.
• The integration of specific captors, enzyme labeling, and real-time sensing facilitates rapid and accurate analysis.
• This automated immunoassay system holds potential for diverse applications in biological and industrial fluid testing.