Development of Enzyme-Linked Immunosorbent Assay for Analysis of Total Aflatoxins Based on Monoclonal Antibody Reactive with Aflatoxins B₁, B₂, G₁ and G₂

Abstract

A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) was developed for the determination of total amount of aflatoxin B 1 , B 2 , G 1 and G 2 (AFB 1 , AFB 2 , AFG 1 and AFG 2 ), using a mouse monoclonal antibody that shows similar reactivity to each of these AFs. The working range of the developed dc-ELISA was 50-230 pg/mL for AFB 1 , 50-270 pg/mL for AFB 2 , 60-390 pg/mL for AFG 1 and 65-700 pg/mL for AFG 2 . The recovery of AFs from spiked roasted peanuts was 98 . Further, when 4 samples actually contaminated with AFB 1 , AFB 2 , AFG 1 and AFG 2 were examined, the results of dc-ELISA were highly correlated with the values assigned by the Food Analysis Performance Assessment Scheme. The developed dc-ELISA appears to be suitable for the determination of total AFs at concentrations around the maximum permitted level (10 μg/kg for all foods) in Japan.

One-Step Ultrasensitive Bioluminescent Enzyme Immunoassay Based on Nanobody/Nanoluciferase Fusion for Detection of Aflatoxin B1 in Cereal

“…Because of the high toxicity and carcinogenicity of aflatoxins, many countries have set up strict maximum limits for AFB 1 or total aflatoxins permissible in food and agriproducts with the range from 2 to 20 μg/kg . Numerous analytical methods have been developed for aflatoxin determination, including chromatography-based instrumental analytical methods and immunoassay-based rapid methods (e.g., ELISA, lateral flow strips, and sensors). − In past decades, numerous antibodies against aflatoxins have been developed by polyclonal and monoclonal techniques and applied in various immunoassay-based rapid methods for the detection of aflatoxins. With the advent of molecular engineering and phage display technology, recombinant antibodies produced in host bacteria or other cell lines have become more useful reagents compared to conventional animal-derived antibodies.…”

A Highly Sensitive and Group-Specific Enzyme-Linked Immunosorbent Assay (ELISA) for the Detection of AFB1 in Agriculture and Aquiculture Products

“…The average IC50 value calculated from the standard curves obtained from six consecutive measurements is 90 pg mL −1 , while the limit of detection (LOD) of the ELISA based on three times the standard deviation (3 × SD) at zero concentration is 18 pg mL −1 . Many mAb (or pAb)-based dc-ELISAs or ic-ELISAs for the detection of AFB1 in different samples have been developed in recent decades (Table 1) [20][21][22][23][24][25][26][27][28][29][30][31][32][33].…”

“…For AFB 2 , although there is no an olefinic bond in the furan ring, due to the high similarity in the molecular structure between AFB 1 and AFB 2 , the CR of the mAb with AFB 2 was as much as 23.6%. Many mAb (or pAb)-based dc-ELISAs or ic-ELISAs for the detection of AFB1 in different samples have been developed in recent decades (Table 1) [20][21][22][23][24][25][26][27][28][29][30][31][32][33].…”

“…In recent decades, many immunoassays, including chemiluminescence immunoassay [16], time-resolved fluorescence immunoassay [17], electrochemical immunosensors [18], and immunochromatographic assay [19], have been reported for the detection of AFB 1 . Furthermore, many direct competitive (dc)-or indirect competitive (ic)-ELISAs based on polyclonal antibodies (pAbs) or monoclonal antibodies (mAbs) for detecting AFB 1 have been developed (Table 1) [20][21][22][23][24][25][26][27][28][29][30][31][32][33]. However, establishing a sensitive immunoassay is mainly dependent upon the quality of antibody.…”

Development of a direct competitive enzyme-linked immunosorbent assay for determination of the fungicide mepanipyrim and its metabolite

“…Such a drastic change had also been found in the ic-ELISA and dc-ELISA for aflatoxin detection. 21,22) MoAbs were immobilized at the well surface in the 96-well microtiter plate in the dc-ELISA but were dissolved in the liquid phase in the ic-ELISA. It seemed that the immobilization might produce minor structural changes to MPP204 that effectively changed its binding ability.…”