scholarly journals Preparation of Monoclonal Antibody and Development of Indirect Competitive Enzyme-Linked Immunosorbent Assay and Fluorescence-Linked Immunosorbent Assay for Detecting 3-Amino-5-methylmorpholino-2-oxazolidinone (AMOZ) in Edible Animal Tissue

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4243
Author(s):  
Yong Xie ◽  
Yarong Wang ◽  
Xueling Yan ◽  
Lu Gan ◽  
Tao Le
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Correlation Coefficients ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Monitoring Methods ◽  
Animal Tissues ◽  
Coefficients Of Variation ◽  
Highly Sensitive ◽  
Ic50 Values

To monitor the illegal used of furaltadone, a highly sensitive indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and fluorescence-linked immunosorbent assay (FLISA) based on a monoclonal antibody (mAb) were developed for the detection of 3-amino-5-methylmorpholino-2-oxazolidinone (AMOZ), the major metabolite of furaltadone in animal tissues. The highly specific mAb, which was very sensitive to a nitrophenyl derivative of AMOZ (2-NP-AMOZ) with IC50 values of 0.11 and 0.09 ng/mL for ic-ELISA and FLISA, respectively, was selected for the development of immunoassays. For both the ic-ELISA and FLISA for AMOZ-spiked experiments, acceptable recovery rates of 81.1–105.3% and coefficients of variation of 4.7–9.8% were obtained. In addition, results from both ic-ELISA and FLISA methods for spiked samples’ data showed excellent correlation coefficients ranging from 0.9652 to 0.9927. Meanwhile, the proposed ic-ELISA and FLISA for thirty spiked samples were confirmed by standard LC-MS/MS with high correlation coefficients of 0.9911 and 0.9921, respectively. These results suggest that the developed ic-ELISA and FLISA are valid and cost-effective tools for high-throughput monitoring methods for AMOZ residues in animal tissues.

An enzyme-linked immunosorbent assay for the detection of Rathayibacter toxicus, the bacterium involved in annual ryegrass toxicity, in hay

2006 ◽  
Vol 57 (7) ◽  
pp. 731 ◽  
Author(s):  
A. M. Masters ◽  
A. R. Gregory ◽  
R. J. Evans ◽  
J. E. Speijers ◽  
S. S. Sutherland
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Specific Antigen ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Annual Ryegrass ◽  
Capture Assay ◽  
Large Numbers ◽  
Soluble Polysaccharide ◽  
Annual Ryegrass Toxicity

An enzyme-linked immunosorbent assay (ELISA) for Rathayibacter toxicus is described. The development of a monoclonal antibody for a specific antigen from R. toxicus and a polyclonal antibody raised against the same R. toxicus preparation enabled a capture assay format. The assay is specific for a soluble polysaccharide produced by the bacterium and was found to be sensitive enough to detect antigen equivalent to less than one gall per kilogram of hay. The applicability of the assay to samples of pasture or hay is demonstrated. Cost-effective testing of large numbers of samples for the presence of R. toxicus is possible with the ELISA. This will assist stockowners, hay producers, and hay exporters in the management of the risk of annual ryegrass toxicity.

scholarly journals Development of a New Monoclonal Antibody against Brevetoxins in Oyster Samples Based on the Indirect Competitive Enzyme-Linked Immunosorbent Assay

Foods ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2398
Author(s):  
Xiya Zhang ◽  
Mingyue Ding ◽  
Chensi Zhang ◽  
Yexuan Mao ◽  
Youyi Wang ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Broad Spectrum ◽  
Limit Of Detection ◽  
Keyhole Limpet Hemocyanin ◽  
Enzyme Linked Immunosorbent Assay ◽  
Neurotoxic Effects ◽  
Rapid Monitoring ◽  
Ic50 Values ◽  
Ester Method

The consumption of shellfish contaminated with brevetoxins, a family of ladder-frame polyether toxins formed during blooms of the marine dinoflagellate Karenia brevis, can cause neurotoxic poisoning, leading to gastroenteritis and neurotoxic effects. To rapidly monitor brevetoxin levels in oysters, we generated a broad-spectrum antibody against brevetoxin 2 (PbTx-2), 1 (PbTx-1), and 3 (PbTx-3) and developed a rapid indirect competitive enzyme-linked immunosorbent assay (icELISA). PbTx-2 was reacted with carboxymethoxylamine hemihydrochloride (CMO) to generate a PbTx-2-CMO hapten and reacted with succinic anhydride (HS) to generate the PbTx-2-HS hapten. These haptens were conjugated to keyhole limpet hemocyanin (KLH) and bovine serum albumin (BSA) to prepare immunogen and coating antigen reagents, respectively, using the active ester method. After immunization and cell fusion, a broad-spectrum monoclonal antibody (mAb) termed mAb 1D3 was prepared. The 50% inhibitory concentration (IC50) values of the icELISA for PbTx-2, PbTx-1, and PbTx-3 were 60.71, 52.61, and 51.83 μg/kg, respectively. Based on the broad-spectrum mAb 1D3, an icELISA was developed to determine brevetoxin levels. Using this approach, the limit of detection (LOD) for brevetoxin was 124.22 μg/kg and recoveries ranged between 89.08% and 115.00%, with a coefficient of variation below 4.25% in oyster samples. These results suggest that our icELISA is a useful tool for the rapid monitoring of brevetoxins in oyster samples.

Detection of Zearalenone By Tandem Immunoaffinity-Enzyme-Linked Immunosorbent Assay and Its Application to Milk

1990 ◽  
Vol 53 (7) ◽  
pp. 577-580 ◽  
Author(s):  
JUAN I. AZCONA ◽  
MOHAMED M. ABOUZIED ◽  
JAMES J. PESTKA
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Monoclonal Antibody ◽  
Initial Sample ◽  
Average Recovery ◽  
Milk Samples ◽  
Coefficients Of Variation ◽  
Direct Elisa ◽  
Commercial Milk

A hybridoma-based method utilizing tandem affinity chromatography and enzyme-linked immunosorbent assay (ELISA) was devised to detect zearalenone. A zearalenone specific monoclonal antibody was attached to Sepharose for initial sample clean-up. Zearalenone was eluted with methanol and then quantified by competitive direct ELISA. Average ELISA recoveries from the column for water spiked with zearalenone at levels of 1, 5, 10, 25, and 50 ng/ml were 107, 86, 95, 95, and 92%, respectively, with a mean recovery of 95%. Mean interwell and interassay coefficients of variation were 9.7 and 8.9%, respectively. Average recovery by the method from milk spiked with zearalenone at levels of 1, 5, 10, 25, and 50 ng/ml was 187, 113, 107, 110, and 112%, respectively, with a mean recovery of 126%. Mean interwell and interassay coefficients of variation were 14.5 and 9.1%, respectively. Zearalenone was not detectable in 12 commercial milk samples assayed by the tandem method.

Comparison of Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay with Thin-Layer Chromatography and Liquid Chromatography for Aflatoxin B1 Determination in Naturally Contaminated Corn and Mixed Feed

1992 ◽  
Vol 75 (2) ◽  
pp. 307-312 ◽  
Author(s):  
Ken-Ichi Hongyo ◽  
Yukikatu Itoh ◽  
Emi Hifumi ◽  
Akira Takeyasu ◽  
Taizo Uda
Keyword(s):  
Monoclonal Antibody ◽  
Liquid Chromatography ◽  
Immunosorbent Assay ◽  
Aflatoxin B1 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mixed Feed ◽  
Elisa System ◽  
Highly Sensitive ◽  
Mixed Feeds ◽  
Layer Chromatography

Abstract A 1-step enzyme-linked Immunosorbent assay (EUSA) method, using a highly sensitive and specific monoclonal antibody to aflatoxin B1 (AFB1), was evaluated by comparison with other methods, Including liquid chromatography (LC) and thinlayer chromatography. The detection limit of the ELISA was as low as 100 pg/assay. AFB1 contents of naturally contaminated corn samples were determined by the 3 methods. The relationships among the methods were Investigated, and good correlations were observed. Mixed feeds were also subjected to AFB1 determination by the 3 methods. For our ELISA system, 3 types of sample preparations were tested. For analysis of mixed feed by ELISA, samples must first be purified by column chromatography. When the relationship between LC and ELISA was also investigated, results were found to have a good correlation coefficient.

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