p-Bromophenol-Enhanced Bienzymatic Chemiluminescence Competitive Immunoassay for Ultrasensitive Determination of Aflatoxin B₁

Abstract

Aflatoxin B1 (AFB1) contamination is one of the most critical global issues in food safety. The high carcinogenic nature necessitates rapid and specific methods for the determination of AFB1 in foodstuffs at ultratrace levels. Here, we report an enhanced bienzymatic chemiluminescence competitive immunoassay for ultrasensitive and high-throughput determination of AFB1. In this assay, protein G was first coated on the wells of a microplate for recognizing the Fc fragment of anti-AFB1 mAbs to reduce the antibody dosage and guarantee high immunological reaction efficiency. The target AFB1 competed with glucose oxidase labeled AFB1 for the limited anti-AFB1 mAbs in the wells of the microplate. p-Bromophenol was employed as an enhancer to obtain intense and long-lasting chemiluminescence. The utilization of an enhancer and bienzymatic catalysts effectively improved the detection sensitivity. The developed method offered a good linearity over 5 orders of magnitude, a detection limit of 5 pg L–1, and a relative standard deviation of 1.9% for AFB1. The application of the developed method to the analysis of grain samples gave quantitative recoveries from 94.0% to 97.0%. The developed method provides a universal platform for high-throughput, ultrasensitive, and high specific detection of pollutants or nutrients in foods.

Review of Research into the Determination of Acrylamide in Foods

“…Immunoassay is one new, rapid, and high-throughput analysis strategy based on the specific combination of antigen (Ag) and antibody (Ab). After nearly 20 years of development, the immunoassay has gradually developed into enzyme-linked immunoassay (ELISA) [74,75], chemiluminescent immunoassay [76,77], fluorescent immunoassay [78,79] and so on, which have been widely used in food analysis, especially in the field of rapid detection. The Ab with specific binding ability is the basis of immunoassay.…”

Vorbeugen oder Heilen – die beispiellose Notwendigkeit von selbstberichtenden Materialien

“…Unglücklicherweise ist die CL-Quantenausbeute von Luminol in polaren aprotischen Lçsungsmitteln (wie DMSO) oder wässrigen Medien eher gering. [123] Daher wurden verschiedene katalytische Systeme entwickelt, die Nanomaterialien, [130,[149][150][151][152] Metallionen [147,148] oder andere Verstärker [153][154][155] füre ine verbesserte CL-Emission enthalten. Da all diese Systeme unter speziellen Nachteilen leiden (u. a. Kosten, Toxizität, Luft-/Feuchtigkeitsempfindlichkeit oder Stabilitätsprobleme), ist die Entwicklung neuer,fortschrittlicher Luminol-CL-Systeme von großer Bedeutung.…”

Luminescence method for detection of aflatoxin B1 using ATP-releasing nucleotides

“…8 Although the above methods are sensitive and accurate, the cost of the detection equipment required is relatively high, and these methods must be operated by professionals under laboratory environment conditions. For the immunoassay method, 9,10 it requires the corresponding antibody of AFB1, however, the antibody needs very harsh storage conditions. The aptamer also has high affinity for the target molecules, but is much easier to label and store.…”