Multiplex highly sensitive immunochromatographic assay based on the use of nonprocessed antisera

Area of Science:

• Analytical Chemistry
• Biotechnology
• Immunotechnology

Background:

• Traditional immunochromatographic assays require extensive antibody purification and conjugation.
• Optimizing antibody and marker concentrations is crucial for assay sensitivity.

Purpose of the Study:

• To propose a novel immunochromatographic multiassay format using native antisera and a universal conjugate.
• To simplify assay development by excluding specific antibody purification and conjugation.
• To enable independent optimization of antibody and marker concentrations for enhanced sensitivity.

Main Methods:

• Development of an immunochromatographic multiassay utilizing native antisera.
• Preparation of a universal conjugate of antispecies antibodies with gold nanoparticles.
• Implementation of the assay for simultaneous detection of atrazine and chlorsulfuron.

Main Results:

• The assay successfully detected atrazine and chlorsulfuron with limits of detection of 0.1 and 0.7 ng/mL, respectively.
• Analysis time was reduced to 20 minutes.
• The assay demonstrated suitability for monitoring herbicides in apple and blackcurrant juices.

Conclusions:

• The proposed assay format significantly simplifies immunochromatographic multiassay development.
• The method offers a low detection limit, high signal intensity, and broad applicability.
• This technique is sensitive, simple, and transferable for detecting various antigens, including herbicides in food matrices.