Abstract
Ochratoxins were important secondary metabolites secreted by fungi, and OTA and OTB are mainly significant mycotoxin, having toxic effects on humans and animals. Therefore, it is important to establish a rapid, sensitive, and precise method for ochratoxins detection and quantification in real samples. In this study, a stable monoclonal antibody (mAb) that recognizing both OTA and OTB toxins was employed for the establishment of indirect competitive ELISA (ic-ELISA), colloidal gold nanoparticles (CGNs), and nanoflowers gold strips (AuNFs) for detection of ochratoxins in real samples. A 6E5 hybridoma cell line stable secreting mAb against both OTA and OTB toxins was obtained by fusion of splenocytes with myeloma SP2/0 cells. The 6E5 mAb had a high affinity (3.7 × 108 L/mol) to OTA, and also showed similar binding activity to OTB. The optimized ic-ELISA resulted in a linear range of 0.06–0.6 ng/mL for ochratoxins (OTA and OTB) detection. The IC50 was 0.2 ng/mL and the limit of detection (LOD) was 0.03 ng/mL. The mean recovery rate from the spiked samples was 89.315 ± 2.257%, with a coefficient variation of 2.182%. The result from lateral flow immunoassays indicated that the LOD of CGNs and AuNFs were 5 and 1 μg/mL, respectively. All these results indicated that the developed ic-ELISA, CGNs, and AuNFs in this study could be used for the analysis of the residual of ochratoxins (OTA and OTB) in food and agricultural products.
Highlights
Mycotoxins are secondary toxic substances produced by fungi in the later process of growth from different types of food, and pose great danger to human’s health, animals, and crops
The titer of anti-Ochratoxin A (OTA)-Bovine serum albumin (BSA) serum isolated from the immunized mice was assayed by indirect ELISA using OTA-Keyhole limpet haemocyanin (KLH) as coating antigen
The supernatants taken from growing hybridoma cells were assessed for specific antibody production against plates coated with OTA-KLH, and the positive hybridomas were screened by indirect ELISA
Summary
Mycotoxins are secondary toxic substances produced by fungi in the later process of growth from different types of food, and pose great danger to human’s health, animals, and crops. Serious diseases such as cancer, tumors, and general weaknesses are often arises when these toxins contaminated foods were consumpted. About 300–400 different types of mycotoxins have been identified so far, and the most famous are Aflatoxin, Ochratoxin, Patulin, Zearalenone, and Trichothecenes among them (Berthiller et al, 2007; Malir et al, 2013). Ochratoxins are a group of mycotoxins produced mainly by Aspergillus species and some Penicillium species (Heussner and Bingle, 2015). Ochratoxin B (OTB) is a non-chlorinated form of ochratoxin A (OTA)
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