Efficient Conjugation of Aflatoxin B1 With Bovine Serum Albumin Applying for Development of Aflatoxin B1 Quick Test

Phong, Truong Quoc; Phạm, Ngọc Thị; Nguyen, Huong; Nguyen, Anh Thi Ngoc

doi:10.15625/2525-2518/56/4a/12887

Cited by 2 publications

(1 citation statement)

References 15 publications

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“…The NMR results showed that the AFB1 antibody and BSA were successfully conjugated to the AuNPs (Figures 1 and 2, Table 1). The most established conjugation of AFB1 to a protein is at the 1-position while using a carboxymethyloxime spacer [37]. However, this position has been taken by the anti-mouse antibody (Figure 2).…”

Section: Confirming Conjugation Of Aunps Using Tlc Gel Electrophoresmentioning

confidence: 99%

Developing Gold Nanoparticles-Conjugated Aflatoxin B1 Antifungal Strips

Sojinrin

Liu

Wang

et al. 2019

IJMS

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Lateral flow immunochromatographic assays are a powerful diagnostic tool for point-of-care tests, based on their simplicity, specificity, and sensitivity. In this study, a rapid and sensitive gold nanoparticle (AuNP) immunochromatographic strip is produced for detecting aflatoxin B1 (AFB1) in suspicious fungi-contaminated food samples. The 10 nm AuNPs were encompassed by bovine serum albumin (BSA) and AFB1 antibody. Thin-layer chromatography, gel electrophoresis and nuclear magnetic resonance spectroscopy were employed for analysing the chemical complexes. Various concentrations of AFB1 antigen (0-16 ng/mL) were tested with AFB1 antibody-BSA-AuNPs (conjugated AuNPs) and then analysed by scanning electron microscopy, ultraviolet-visible spectroscopy, and Zetasizer. The results showed that the AFB1 antibody was coupled to BSA by the N-hydroxysuccinimide ester method. The AuNPs application has the potential to contribute to AFB1 detection by monitoring a visible colour change from red to purple-blue, with a detection limit of 2 ng/mL in a 96-well plate. The lateral flow immunochromatographic strip tests are rapid, taking less than 10 min., and they have a detection capacity of 10 ng/g. The smartphone analysis of strips provided the results in 3 s, with a detection limit of 0.3 ng/g for AFB1 when the concentration was below 10 ng/g. Excellent agreement was found with AFB1 determination by high-performance liquid chromatography in the determination of AFB1 among 20 samples of peanuts, corn, rice, and bread.for Aflatoixin B1 (AFB1) [8]. The European Economic Community (EEC) has established permitted food contamination limits of 2 µg/kg for AFB1 and 4 µg/kg for the total concentration of the four AFLs since 1 February 1999 [9]. Therefore, it is necessary to develop strategies for achieving the limits of AFL contamination and reducing AFL exposure in vulnerable populations [10].Thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) are the most popular techniques for detecting AFLs. However, these methods require extensive sample preparation, expensive instruments, and operation by skilled professionals. Alternatively, the enzyme-linked immunosorbent assay (ELISA) has been successfully developed for AFLs [11], but ELISA also needs incubation and washing steps, and application is mainly confined to laboratories. Lateral flow immunochromatographic/immunoassay strips (LFIAs) have received increasing attention for qualitative and quantitative analysis in different scientific sectors [12], including food safety, environmental monitoring, and precision medicine [12,13]. In 2005, Delmulle et al. [14] developed an LFIA for the detection of aflatoxin B1 (AFB1) in pig feed. Liao and Li [15] have made significant effort to investigate the effect of the core-shell silver-gold nanocomposites on the properties of LFIAs. However, this detection can only provide either qualitative (positive or negative) or semi-quantitative information on analyte concentration, and thereby does not satisfy the requirements for...

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Section: Confirming Conjugation Of Aunps Using Tlc Gel Electrophoresmentioning

confidence: 99%

Developing Gold Nanoparticles-Conjugated Aflatoxin B1 Antifungal Strips

Sojinrin

Liu

Wang

et al. 2019

IJMS

View full text Add to dashboard Cite

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Antibody conjugated magnetic nanoparticle based colorimetric assay for the detection and quantification of aflatoxin B1 in wheat grains

Kumari

Jaiswal

Chowdhury

et al. 2022

WMJ

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Aflatoxin B1 (AFB1) is a most potent carcinogenic secondary metabolite produced by Aspergillus flavus. As a food safety concern, development of a rapid, cost effective, sensitive and easy to use method for the detection of aflatoxin is of prime requirement. In this study, AFB1 was conjugated with bovine serum albumin (BSA), and AFB1-BSA conjugate was purified by HPLC. Purification was confirmed by UV-Vis spectroscopy, FTIR and MALDI-TOF mass spectrometry. The polyclonal antibody was raised against AFB1-BSA conjugate in rabbit and purified by protein A sepharose and BSA sepharose affinity columns. Iron oxide nanoparticles (MNPs) were synthesised by co-precipitation method and their surface was functionalised with (3-aminopropyl) triethoxysilane (APTES). Size of APTES conjugated MNPs was determined by electron microscopy, and characterised by several biophysical techniques. The purified anti-AFB1 antibody was conjugated with surface functionalised MNPs and the conjugation was confirmed by determining the sizes of free and antibody conjugated MNPs by field emission scanning electron microscope where increase of particle sizes from 10-20 to 40-50 nm was observed due to antibody conjugation. Anti-AFB1 antibody conjugated MNPs were used for capturing AFB1 from the aflatoxin spiked wheat grains with a recovery percentage of more than 80% and used effectively five times. The captured AFB1 was then quantified by a sensitive colorimetric assay where colourless AFB1 was first converted into coumaric acid by NaOH. Subsequently, coumaric acid reacted with 2,6-dibromoquinone-4-chloroimide (DBQC) to a green-coloured indophenol product which was quantified spectrophotometrically. AFB1 contamination as low as 2 μg/kg in wheat grains was detected by the developed technique suggesting its potential application for both qualitative and quantitative analysis of aflatoxins present in feed and food materials.

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Efficient Conjugation of Aflatoxin B1 With Bovine Serum Albumin Applying for Development of Aflatoxin B1 Quick Test

Cited by 2 publications

References 15 publications

Developing Gold Nanoparticles-Conjugated Aflatoxin B1 Antifungal Strips

Developing Gold Nanoparticles-Conjugated Aflatoxin B1 Antifungal Strips

Antibody conjugated magnetic nanoparticle based colorimetric assay for the detection and quantification of aflatoxin B1 in wheat grains

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