Highly efficient dual-mode detection of AFB1 based on the inner filter effect: Donor-acceptor selection and
Jincheng Xiong1, Boyan Sun2, Shuai Zhang2
1National Key Laboratory of Veterinary Public Health Security, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China; Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, China Agricultural University, Beijing, 100193, China; Guangdong Provincial Key Laboratory of Advanced Biomaterials, Shenzhen Key Laboratory of Smart Healthcare Engineering, Department of Biomedical Engineering, Southern University of Science and Technology, Shenzhen, Guangdong, 518055, China.
This study developed a sensitive fluorescence immunoassay for Aflatoxin B1 detection using gold nanoclusters and the inner filter effect. The method offers improved sensitivity for food safety monitoring.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Materials Science
Background:
- Inner filter effect (IFE) offers potential for fluorescence immunoassays but selecting optimal donors is challenging.
- Aflatoxin B1 (AFB1) is a toxic mycotoxin found in agricultural products, necessitating sensitive detection methods for food safety.
- Developing convenient and sensitive approaches for AFB1 detection is crucial for public health.
Purpose of the Study:
- To explore tunable multicolor fluorescence of gold nanoclusters (AuNCs) for maximizing IFE efficiency with p-nitrophenol (PNP).
- To develop a sensitive dual-mode immunoassay for Aflatoxin B1 (AFB1) monitoring by integrating IFE, enzymatic reactions, and antibody-antigen recognition.
- To establish a screening procedure for selecting optimal donor-acceptor pairs in IFE analysis.
Main Methods:
- Utilized alkaline phosphatase (ALP) to catalyze the hydrolysis of p-nitrophenylphosphate to PNP, serving as a model reaction.
- Selected green-emitting AuNCs as optimal donors for IFE due to spectral overlap, high photoluminescence, and system adaptability.
- Validated the IFE mechanism between PNP and AuNCs through UV-Vis absorption, zeta potential, and fluorescence lifetime measurements.
- Integrated antibody-antigen recognition and enzymatic reaction with AuNCs for a dual-mode immunoassay for AFB1 detection.
Main Results:
- Identified green-emitting AuNCs as the optimal donor, achieving a 22-fold sensitivity increase for ALP detection compared to colorimetric methods.
- Developed a fluorometric immunoassay for AFB1 with a sensitivity of 0.06 ng/mL, a 3.5-fold improvement over ELISA.
- Confirmed the practicability and applicability of the developed method in real samples like tap water, corn, wheat, and peanuts.
Conclusions:
- Provided a straightforward method for selecting optimal donor-acceptor pairs in IFE analysis.
- Demonstrated that integrating IFE-based signal conversion into immunoassays can simplify amplification, reduce errors, and enhance signal types.
- Proposed the IFE strategy as a versatile approach for monitoring trace levels of various contaminants.
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