Sensitive competitive immunoassay of multiple mycotoxins with non-fouling antigen microarray

Area of Science:

• Food Safety
• Analytical Chemistry
• Biomaterials

Background:

• Mycotoxins pose significant food safety risks due to their carcinogenicity and toxicity.
• Sensitive and high-throughput detection methods are crucial for monitoring these contaminants.
• Existing methods may lack the sensitivity or throughput required for comprehensive food safety analysis.

Purpose of the Study:

• To develop a sensitive and high-throughput microarray for detecting multiple mycotoxins.
• To utilize a non-fouling polymer brush, poly[(ethylene glycol) methacrylate-co-glycidyl methacrylate] (POEGMA-co-GMA), for improved immunoassay performance.
• To establish a powerful tool for rapid screening of toxins in food and environmental samples.

Main Methods:

• Fabrication of a fluorescent competitive immunoassay microarray using a POEGMA-co-GMA polymer brush.
• Immobilization of target mycotoxins (aflatoxin B1, ochratoxin A, zearalenone) as template antigens.
• Evaluation of microarray performance, including dynamic range and detection limits.
• Comparison with epoxy-functionalized microarrays and enzyme-linked immunosorbent assay (ELISA).

Main Results:

• The POEGMA-co-GMA brush provided uniform protein loading and high resistance to nonspecific absorption.
• The optimal microarray achieved wide dynamic ranges across three orders of magnitude.
• Low detection limits were obtained: 4 pg mL(-1) for aflatoxin B1, 4 pg mL(-1) for ochratoxin A, and 3 pg mL(-1) for zearalenone.
• Performance surpassed epoxy-functionalized microarrays and was comparable or superior to ELISA.

Conclusions:

• The developed polymer brush-based microarray is a highly sensitive and efficient tool for multiplexed mycotoxin detection.
• This technology offers a significant advancement for high-throughput screening in food safety and environmental monitoring.
• The non-fouling properties of the polymer brush are key to the enhanced performance of the immunoassay.