Preparation of monoclonal antibody and development of an indirect competitive enzyme-linked immunosorbent assay for ornidazole detection

Abuse of pyrimethanil in crops to cause deleterious effects on humans has been of great concern. In this study, functionalized derivatives of pyrimethanil have been synthesized and conjugated to carrier proteins, and highly specific and sensitive monoclonal antibodies (mAb) have been produced. Based on the mAb 1F05, an ic-ELISA was developed and characterized for the analysis of pyrimethanil residues. The standard curves based on the pyrimethanil matrix calibration ranged from 0.03 to 3 ng/mL, with a 50% inhibitory concentration of 0.25 ng/mL. The limits of detection were 0.02-0.04 μg/kg in various matrices. The recoveries for pyrimethanil ranged from 88.6% to 102.3%, with coefficient of variation below 11%. A good correlation (R 2 > 0.9825) between the ic-ELISA and HPLC-MS/MS showed the reliability of the developed ic-ELISA. The proposed ic-ELISA was demonstrated to be a simple, rapid, and sensitive tool to monitor the residues of pyrimethanil in fruits and vegetables.

Section: Hapten Design and Antigen Preparationmentioning

confidence: 56%

Development a monoclonal antibody-based enzyme-linked immunosorbent assay for screening pyrimethanil in fruits and vegetables

Cao¹,

Shi

et al. 2019

“…In the final step, 50 μL of 2 M sulfuric acid was added to terminate the enzymatic reaction and it was then tested at 450 nm using a microplate reader. Each test was repeated four times Zhao et al, 2017).…”

Section: Ic-elisa Proceduresmentioning

confidence: 99%

Gold nanoparticle-based lateral flow strips for rapid and sensitive detection of Virginiamycin M1

Kuang

et al. 2020

In this study, a sensitive and specific monoclonal antibody against VIR M1 was produced based on a new hapten. The antibody had a linear detection range of 3.77-65.51 ng/mL with an IC 50 of 15.26 ng/mL and good specificity for VIR M1. Gold nanoparticle (GNP)-based lateral flow strips were assembled for rapid screening of VIR M1 in feed samples. When evaluated with the naked eye, the visual limit of detection (LOD) in feed samples was 25 μg/kg, and the cutoff value was 200 μg/kg. Therefore, this GNP-based lateral flow strip assay can be used for on-site detection and rapid preliminary screening of VIR M1 in feed samples.

“…Due to the presence of carboxyl groups in haptens, we chose the active ester method (Chen et al, 2017a;Liu et al, 2018;Zhao et al, 2017) to couple the small molecules to the carrier proteins. Briefly, a mixture of AMA-hapten (10.5 mg, 0.04 mmol) and NHS (13.7 mg, 0.12 mmol) was dissolved in 2 mL of DMF.…”

Section: Preparation Of the Haptens And Antigensmentioning

confidence: 99%

“…After three days, cell fusion between splenocytes from the immunized mouse and SP2/0 myeloma cells was carried out under sterile conditions (Xie et al, 2018;Zhao et al, 2017). The culture supernatants were collected and monitored by ELISA and ic-ELISA after 7 days.…”

Section: Preparation Of the Mabmentioning

confidence: 99%

A sensitive lateral flow immunoassay for the multiple residues of five adamantanes

Gao

Wang

et al. 2019

Adamantanes (ADAs) are tricyclic decane saturated derivatives, which are used in the protection against the influenza virus. However, their abuse in animal husbandry has affected human health. To help prevent drug resistance and protect food safety, a rapid, portable, sensitive, and reliable method for onsite detection of ADAs was developed. Herein, we have produced a broadspecificity anti-ADAs monoclonal antibody (mAb), the subclass of which is IgG 2b , and its affinity constant is 7.25 × 10 9 L/mol. From this, an antibody-based lateral flow immunoassay (LFIA) was developed for the rapid detection of five ADAs in food samples. When viewed with the naked eye, the LFIA had a visual detection limit of 0.1, 0.1, 1, 5, and 10 μg/kg and a cutoff value of 2.5, 2.5, 10, 100, and 100 μg/kg for somantadine, rimantadine, amantadine, 1-adamantanemethanol, 1-adamantyl methyl ketone respectively in chicken samples. The test results for the LFIA were very similar to that seen when using an enzyme linked immunosorbent assay (ELISA). Therefore, this paper-based LFIA provides a useful tool for rapid, onsite detection of ADAs in biological samples.