Preparation of a monoclonal antibody against testosterone and its use in development of an immunochromatographic assay

Self Cite

In this study, we prepared a monoclonal antibody against pyrimethanil, for which we first derived its hapten according to the structural formula of pyrimethanil; we used indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) to screen positive mice for cell fusion; finally, we developed five cell lines (1A5, 1E7, 3A6, 3H12 and 5D4) that produced monoclonal antibodies to pyrimethanil. The monoclonal antibody produced by each cell was detected by ic-ELISA, and the monoclonal antibody produced by the 1E7 cell strain was found to be the most sensitive to pyrimethanil. The IC50 value of this antibody was 4 ng/ml. After using this monoclonal antibody, i.e. 1E7, to perform immunocolloidal gold strip treatment on cucumber samples, the PBS and the cucumber samples cutoff value of Immunochromatographic strips (ICS) was found to be 50 ng/ml; We conclude that ICS can rapidly detect pyrimethanil in cucumber samples.

Section: Preparation Of Monoclonal Antibodies (Mabs) Against Pyrmentioning

confidence: 99%

“…We then dissolved the precipitate in 1 mL ultrapure water and centrifuged it. Finally, were suspended the gold-labelled anti-PYR mAb in borate buffer (BB) (0.002M BB, 1% sucrose, and 0.01% Tween-20, pH 7.2, w/v) and stored at 4°C (Wang et al, 2016).…”

Section: Preparation Of Gold-labelled Anti-pyr Mabmentioning

confidence: 99%

Development of immunocolloidal strip for rapid detection of pyrimethanil

Chen

et al. 2019

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“…The common techniques used to detect defective foods are liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry. Nevertheless, these methods are not suitable for on-site analysis (Wang et al, 2016;Xu et al, 2006); moreover, they are expensive and require more skilled personnel to operate them. Therefore, rapid, sensitive, inexpensive and accurate simple and time-saving methods such as Lateral immunochromatographic strips and indirect competitive enzyme-linked immunosorbent assay (ic-ELISA), which require less knowledge for operation, must be developed complementary to conventional instrumental techniques Tochi et al, 2015;Xu et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Development of an immunochromatographic assay for hexestrol and diethylstilbestrol residues in milk

Mukunzi

Tochi

Isanga

et al. 2016

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Hexestrol (HES) and Diethylstilbestrol (DES) are synthetic hormones, which have been found abuse use in animal-origin food production. We developed an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and immune-chromatographic strip to detect these compounds based on monoclonal antibody. Under optimized conditions, the half-inhibition concentration (IC 50 ) values of ic-ELISA were 0.15 µgL −1 and 0.23 µgL −1 for HES and DES, respectively. Spiked milk samples were analyzed. The recovery of both synthetic hormones in the milk samples was 60.48-102.19% (HES) and 89.34-100.16% (DES). In immunechromatographic assay, the visual cutoff values at 0.5 and 1.0 µgL −1 respectively for HES and DES, which allow us to detect milk samples of a concentration low to 10 µgL −1 for HES and 15 µgL −1 for DES. ARTICLE HISTORY

“…The enzyme-linked immunosorbent assay (ELISA) and the lateral flow strip test allow the semi-quantitative analysis of various drug residues and have the advantages of simplicity, rapidity, specificity, convenience, high sensitivity, and cost effectiveness, and the samples need only be pretreated simply before detection (Gu, Liu, Song, Kuang, & Xu, 2016;Peng, Song, Liu, Kuang, & Xu, 2016;Wang et al, 2016;Yu, Liu, Song, Kuang, & Xu, 2016).…”

Section: Introductionmentioning

confidence: 99%

Development of a monoclonal antibody assay and a lateral flow strip test for the detection of paromomycin residues in food matrices

Isanga

Mukunzi

Suryoprabowo

et al. 2017

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A monoclonal antibody (MAb) was produced against paromomycin and used in the development of an immunoassay and a lateral flow strip test for the detection of paromomycin residues in food matrices. The MAb had 25.1% cross-reactivity with neomycin, but was insensitive to other aminoglycosides. Tested under optimized conditions in 0.01 M phosphate-buffered saline, the half maximum inhibitory concentration (IC 50 ) of the MAb was 0.61 ng/ml for paromomycin and 2.43 ng/ml for neomycin, with results obtained within 90 min. The mean recoveries from spiked food matrices were within the range of 64.56-105.85% for paromomycin and 54.08-100.55% for neomycin. The strip test results for different food matrices were obtained within 5 min and showed visual detection limits of 2.5-20 ng/ml (µg/kg) for paromomycin and 10-30 ng/ml for neomycin. Therefore, the developed immunoassay and strip test can be used in food analysis for routinely monitoring not only paromomycin but also neomycin residues. ARTICLE HISTORY