Immunoanalytical methods for ochratoxin A monitoring in wine and must based on innovative immunoreagents

López-Puertollano, Daniel; Agulló, Consuelo; Mercader, Josep V.; Abad‐Somovilla, Antonio; Abad‐Fuentes, Antonio

doi:10.1016/j.foodchem.2020.128828

Cited by 12 publications

(6 citation statements)

References 37 publications

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“…It can be seen from Figure 3 that the average IC50 value calculated from standard curves obtained on six consecutive days was 34.8 pg mL −1 , while the limit of detection (LOD) of the ELISA based on three times the standard deviation (3×SD) at zero concentration was found to be 1.5 pg mL −1 . In the last decades, many ELISAs for the detection of OTA in food samples based on pAb or mAb have been developed [23][24][25][26][27][28][29][30][31][32][33][34][35][36]. As shown in Table 1, the IC50 (or LOD) value achieved in our ELISA is about 1.53~147 times lower than those obtained in other ELISAs, which clearly demonstrates the high sensitivity of our ELISA.…”

Section: Sensitivity Of the Elisa For Otamentioning

confidence: 63%

“…In the last decades, many immunoassays using different signal readouts, such as chemiluminescence [20], time-resolved fluorescence [21], electrochemical [22], etc., have reported the detection of OTA. Also, many polyclonal antibody (pAb)or monoclonal antibody (mAb)-based indirect competitive ELISAs (ic-ELISA) and direct competitive ELISAs (dc-ELISA) have been developed for the detection OTA in food samples (Table 1) [23][24][25][26][27][28][29][30][31][32][33][34][35][36]. Usually, for the developed immunoassays, the first purpose is to achieve the high sensitivity of the assay for the target analyte.…”

Section: Introductionmentioning

confidence: 99%

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An Extremely Highly Sensitive ELISA in pg mL−1 Level Based on a Newly Produced Monoclonal Antibody for the Detection of Ochratoxin A in Food Samples

Ren,

Tian,

Wang

et al. 2023

Molecules

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In this study, an extremely highly sensitive enzyme-linked immunosorbent assay (ELISA) based on a newly produced monoclonal antibody (mAb) for the detection of ochratoxin A (OTA) in food samples was developed. OTA-Bovine serum albumin (BSA) conjugate was prepared and used as the immunogen for the production of the mAb. Among four hybridoma clones (8B10, 5C2, 9B7, and 5E11), the antibody from 8B10 displayed the highest affinity recognition for OTA. Based on the mAb (8B10), the IC50 and LOD of the ELISA for OTA were 34.8 pg mL−1 and 1.5 pg mL−1, respectively, which was 1.53~147 times lower than those in published ELISAs, indicating the ultra-sensitivity of our assay. There was no cross-reactivity of the mAb with the other four mycotoxins (AFB1, ZEN, DON, and T-2). Due to the high similarity in molecular structures among OTA, ochratoxin B (OTB), and ochratoxin C (OTC), the CR values of the mAb with OTB and OTC were 96.67% and 22.02%, respectively. Taking this advantage, the ELISA may be able to evaluate total ochratoxin levels in food samples. The recoveries of the ELISA for OTA in spiked samples (corn, wheat, and feed) were 96.5–110.8%, 89.5–94.4%, and 91.8–113.3%; and the RSDs were 5.2–13.6%, 8.2–13.0%, and 7.7–13.7% (n = 3), respectively. The spiked food samples (corn) were measured by ELISA and HPLC-FLD simultaneously. A good correlation between ELISA (x) and HPLC-FLD (y) with the linear regression equation of y = 0.918x − 0.034 (R2 = 0.985, n = 5) was obtained. These results demonstrated that the newly produced mAb-based ELISA was a feasible and ultra-sensitive analytical method for the detection of OTA in food samples.

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Section: Sensitivity Of the Elisa For Otamentioning

confidence: 63%

Section: Introductionmentioning

confidence: 99%

An Extremely Highly Sensitive ELISA in pg mL−1 Level Based on a Newly Produced Monoclonal Antibody for the Detection of Ochratoxin A in Food Samples

Ren,

Tian,

Wang

et al. 2023

Molecules

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show abstract

“…Different mycotoxins have been found not only in fruits and vegetables but also in their derivative products [ 8 , 9 , 10 , 11 , 12 ]. The mycotoxins commonly found in fruits and vegetables can be divided into four major categories: PAT, TCs, OTA and ATs.…”

Section: Occurrence Contamination and Toxicity Of Mycotoxinsmentioning

confidence: 99%

Contamination, Detection and Control of Mycotoxins in Fruits and Vegetables

Nan

Xue

2022

Toxins

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Mycotoxins are secondary metabolites produced by pathogenic fungi that colonize fruits and vegetables either during harvesting or during storage. Mycotoxin contamination in fruits and vegetables has been a major problem worldwide, which poses a serious threat to human and animal health through the food chain. This review systematically describes the major mycotoxigenic fungi and the produced mycotoxins in fruits and vegetables, analyzes recent mycotoxin detection technologies including chromatography coupled with detector (i.e., mass, ultraviolet, fluorescence, etc.) technology, electrochemical biosensors technology and immunological techniques, as well as summarizes the degradation and detoxification technologies of mycotoxins in fruits and vegetables, including physical, chemical and biological methods. The future prospect is also proposed to provide an overview and suggestions for future mycotoxin research directions.

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“…In the past decades, numerous analytical techniques have been developed to detect OTA, including high performance liquid chromatography (HPLC) [ 12 ], liquid chromatography tandem mass spectrometry (LC-MS/MS) [ 13 ], enzyme-linked immunosorbent assay (ELISA) [ 14 ], lateral flow immunoassay (LFIA) [ 15 ] and biosensors [ 16 ]. Among these, ELISA is the most commonly used rapid screening method due to its high throughput and simple operations [ 17 ].…”

Section: Introductionmentioning

confidence: 99%

Nanobody-Nanoluciferase Fusion Protein-Enabled Immunoassay for Ochratoxin A in Coffee with Enhanced Specificity and Sensitivity

Bao

Liu

Liao

et al. 2022

Toxins

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Ochratoxin A (OTA), one of the best-known mycotoxins, causes problems concerning food safety with potential toxic effects in humans and animals. So, it is crucial to develop simple and sensitive methods for the detection of OTA. Herein, a nanoluciferase–nanobody fusion protein (Nb28-Nluc)-retaining antibody recognition and enzymatic activity was first prepared, which was then applied as a bifunctional tracer to construct a one-step bioluminescent enzyme-linked immunosorbent assay (BLEIA) for OTA in coffee samples. On the basis of Nb28-Nluc, the BLEIA can be completed with a one-step incubation and detection, with only a substrate replacement from 3,3′,5,5′-tetramethylbenzidine (TMB) to a Nluc assay reagent (Furimazine). Under the optimal experimental conditions, the proposed one-step BLEIA achieved a detection limit of 3.7 ng/mL (IC10) within 3 h. Moreover, the BLEIA method showed good repeatability and accuracy in the spike recovery experiments with recoveries of 83.88% to 120.23% and relative standard deviations (RSDs) of 5.2% to 24.7%, respectively. Particularly, the BLEIA displayed superior performances, such as fewer operations and more rapid and sensitive detection as compared with Nb28-based enzyme-linked immunosorbent assay. Therefore, the proposed one-step BLEIA has great potential for the sensitive and accurate screening of OTA in food samples.

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Immunoanalytical methods for ochratoxin A monitoring in wine and must based on innovative immunoreagents

Cited by 12 publications

References 37 publications

An Extremely Highly Sensitive ELISA in pg mL−1 Level Based on a Newly Produced Monoclonal Antibody for the Detection of Ochratoxin A in Food Samples

An Extremely Highly Sensitive ELISA in pg mL−1 Level Based on a Newly Produced Monoclonal Antibody for the Detection of Ochratoxin A in Food Samples

Contamination, Detection and Control of Mycotoxins in Fruits and Vegetables

Nanobody-Nanoluciferase Fusion Protein-Enabled Immunoassay for Ochratoxin A in Coffee with Enhanced Specificity and Sensitivity

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