Development of ELISA and Lateral Flow Immunoassays for Ochratoxins (OTA and OTB) Detection Based on Monoclonal Antibody

Frontiers in Cellular and Infection Microbiology

Shihua Wang²  Rongzhi Wang²  Sumei Ling²  Shiwei Xiao²  Ke Wang³  Mohamed Hassan Fadlalla³  Jun Yuan³  Xiulan Li³  Shuqin Tang³  Hoyda Elsir³ 

[1] Fujian Key Laboratory of Pathogenic Fungi and Mycotoxins, Fujian Agriculture and Forestry University, Fuzhou, China;Key Laboratory of Biopesticide and Chemical Biology of the Education Ministry, Fujian Agriculture and Forestry University, Fuzhou, China;School of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou, China;

关键词: ochratoxins;    monoclonal antibody;    hybridoma technology;    ic-ELISA;    colloidal gold strip;    nanoflowers gold strip;   

DOI  :  10.3389/fcimb.2020.00080

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown