Development of Enzyme Immunoassay for Captan and Its Degradation Product Tetrahydrophthalimide in Foods

1993 ◽  
Vol 76 (2) ◽  
pp. 381-386 ◽  
Author(s):  
W Harvey Newsome ◽  
Jupiter M Yeung ◽  
Peter G Collins
Keyword(s):  
Detection Limit ◽  
Human Serum Albumin ◽  
Degradation Product ◽  
Protein Concentration ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Compound ◽  
Coefficients Of Variation ◽  
Spacer Arm ◽  
Related Compounds

Abstract A simple, sensitive, and precise enzyme-linked immunosorbent assay (ELISA) is described for the quantitation of captan as its degradation product tetrahydrophthalimide (THPI) in foods using polyclonal antibodies. Three hapten analogues of THPI with different alky I spacer arm lengths were synthesized. Immunogens and coating proteins were prepared by coupling these haptens to human serum albumin and ovalbumin, respectively. A 5-carbon spacer arm appeared to be optimum for the production of antibodies. Heterologous coating proteins did not improve the sensitivity, but reduction of homologous coating protein concentration did improve the sensitivity, resulting in a concentration of test compound required to inhibit binding by 50% of 15.5 ng/mL The antiserum is specific for captan, captafol, and THPI, but not other structurally related compounds. The minimum detection limit was 1 ng/mL; the linearity was 1-200 ng/mL. The overall recoveries of captan and THPI from 11 commodities spiked at 4 levels were 92 and 100%, respectively. The intra-assay and interassay coefficients of variation were 9.1 and 16.8% for apple blanks and 5.9 and 4.2% for apple spiked with 3 ppm THPI, respectively. The ELISA described is suitable for measuring captan and THPI at levels comparable to those typically found in fruit.

scholarly journals Determination of Egg Proteins in Food Products by Enzyme Immunoassay

2000 ◽  
Vol 83 (1) ◽  
pp. 139-143 ◽  
Author(s):  
Jupiter M Yeung ◽  
W Harvey Newsome ◽  
Michael A Abbott
Keyword(s):  
Detection Limit ◽  
Enzyme Immunoassay ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antigen Binding ◽  
Food Ingredients ◽  
Coefficients Of Variation ◽  
Egg Products ◽  
Whole Egg

Abstract An enzyme-linked immunosorbent assay (ELISA) was developed to determine the presence of egg proteins in foods. The polyclonal antibodies developed were specific to whole egg proteins and did not cross-react with any of the 38 nuts, legumes, or other common food ingredients tested. The concentrations of egg proteins that will inhibit 50% of antibody–antigen binding, IC50, were 3–7 ng/mL, and the linear range was 0.5–62.5 ng/mL. The detection limit was 0.2 ppm for various foods. Recoveries ranged from 67 to 96%. The intra- and inter-assay coefficients of variation in this procedure were 10–13% for ice cream spiked at 0.8 and 1.6 ppm. The ELISA has been applied to ice creams, noodles, pasta, and breads. Egg proteins were identified in all declared egg products, and no false positives were found.

Production and Characterization of Antibodies against Fumonisin B1

1996 ◽  
Vol 59 (9) ◽  
pp. 992-997 ◽  
Author(s):  
FENG-YIR YU ◽  
FUN S. CHU
Keyword(s):  
Detection Limit ◽  
Polyclonal Antibodies ◽  
Fumonisin B1 ◽  
Correlation Coefficients ◽  
Keyhole Limpet Hemocyanin ◽  
Hplc Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Indirect Competitive Elisa ◽  
Antibody Titers

Polyclonal antibodies against fumonisin B1 (FmB1) were produced in rabbits after immunizing the animals with either FmBl-keyhole limpet hemocyanin (KLH) or FmB1 bovine serum albumin (BSA). A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) and an indirect competitive ELISA (idc-ELISA) were used for the characterization of the antibodies and for analysis of the toxin in corn samples. The antibody titers in the serum of rabbits immunized with FmBl-KLH were considerably higher than in those immunized with FmBl-BSA. The antibodies from the rabbits immunized with FmBl-KLH were further characterized. The concentrations causing 50% inhibition of binding of FmB1-horseradish peroxidase (HRP) to the antibodies by FmB1, FmB2 and FmB3 in the ELISA were found to be 0.45, 0.72, and 25 ng/ml, respectively. The detection limit of FmBl, based on 95% confidence at 5% of inhibition of binding of FmBl-HRP conjugate, in buffer of the dc-ELISA was found to be 0.05 ng/ml. In the presence of a matrix such as corn, the detection limit was less than 50 ppb. The overall analytical recoveries of FmBl (50 to 1,000 ng/g) added to the ground corn and then extracted with CH3CN/H2O (1/1, vol/vol) with cleanup and without cleanup in the dc ELISA were found to be 70.5 and 85.9%, respectively. A good correlation was found between the FmBl levels in 2 starch and 10 naturally contaminated corn samples analyzed by the dc-ELISA and the high-pressure liquid chromatography (HPLC) method. The correlation coefficients between ELISA and HPLC were found to be 0.955 (y [ELISA] = 1.3 1x [HPLC] + 77 ppb; P < 0.001) and 0.811 (y = 1.13x + 34 ppb; P < 0.01) for the sample without and with cleanup treatment, respectively.

Development of an Analytical Protocol for the Determination of Polybrominated Biphenyls in Water by Enzyme-Linked Immunosorbent Assay

2011 ◽  
Vol 347-353 ◽  
pp. 1537-1541 ◽  
Author(s):  
Han Yu Chen ◽  
Hui Sheng Zhuang
Keyword(s):  
Detection Limit ◽  
Quantitative Determination ◽  
Incubation Time ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibody Concentration ◽  
Polybrominated Biphenyls ◽  
Analytical Protocol ◽  
Optimized Conditions

A novel immunoassay has been developed for the quantitative determination of polybrominated biphenyls (PBBs) using indirect competitive format. A new method was developed to synthesize PBBs congener (PCB15) hapten and its artificial immunogen, then the polyclonal antibodies. The assay was optimized concerning the coating conjugate and antibody concentration, incubation time and temperature, the tolerance to organic solvents and so on. Under optimized conditions, PBB15 can be determined in the concentration range of 0.01-100 μg L-1 with a detection limit of 0.02 μg L-1. The cross-reactivities of the assays were below 8%. While water samples could be analyzed directly.

scholarly journals Development of new immunoanalytical test systems for diagnostics of potato blackleg caused by Dickeya spp. bacteria

2021 ◽  
Vol 16 (3) ◽  
pp. 198-214
Author(s):  
Shyatesa Razo ◽  
Pavel A. Galushka ◽  
Yuri A. Varitsev ◽  
Anatoly V. Zherdev ◽  
Irina V. Safenkova ◽  
...  
Keyword(s):  
Detection Limit ◽  
Polyclonal Antibodies ◽  
Test System ◽  
Diagnostic Tools ◽  
Enzyme Linked Immunosorbent Assay ◽  
Potato Seed ◽  
Potato Blackleg ◽  
Bacterial Diseases ◽  
High Affinity ◽  
Test Systems

Potato blackleg caused by Dickeya spp. bacteria is one of the most important bacterial diseases of potatoes. The rapid spread of this disease in the territory of Russia requires new effective diagnostic tools for the timely detection of infection. To solve this problem, antisera specific to Dickeya spp. were obtained. Polyclonal antibodies isolated from antisera have shown high affinity for the main species of Dickeya spp. ( D. solani, D. dianthicola, D. chrysanthemi, D. dadantii, D. paradisiaca ). Enzyme linked immunosorbent assay (ELISA) and lateral flow immunoassay (LFIA) test systems have been developed based on specific and high affinity antibodies that were obtained. For ELISA, the detection limit was 0.8 105 cells/mL for D. solani and 2 104 cells/mL for D. dianthicola . For LFIA, suitable for use in non-laboratory conditions, the detection limit of D. solani was 2 105 cells/mL and the analysis time was 15 minutes. When testing potato seed material, LFIA test system confirmed positive results of ELISA determination in 75 % of samples, and negative - in 100 % of samples.

scholarly journals How to Improve Sensitivity of Sandwich Lateral Flow Immunoassay for Corpuscular Antigens on the Example of Potato Virus Y?

Sensors ◽  
2018 ◽  
Vol 18 (11) ◽  
pp. 3975 ◽  
Author(s):  
Shyatesa Razo ◽  
Vasily Panferov ◽  
Irina Safenkova ◽  
Yuri Varitsev ◽  
Anatoly Zherdev ◽  
...  
Keyword(s):  
Detection Limit ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Polyclonal Antibodies ◽  
Test Strip ◽  
Lateral Flow ◽  
Lateral Flow Immunoassay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Quantitative Results ◽  
Solanaceae Family

A simple approach was proposed to decrease the detection limit of sandwich lateral flow immunoassay (LFIA) by changing the conditions for binding between a polyvalent antigen and a conjugate of gold nanoparticles (GNPs) with antibodies. In this study, the potato virus Y (PVY) was used as the polyvalent antigen, which affects economically important plants in the Solanaceae family. The obtained polyclonal antibodies that are specific to PVY were characterized using a sandwich enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR). For LFIA, the antibodies were conjugated with GNPs with a diameter of 17.4 ± 1.0 nm. We conducted LFIAs using GNP conjugates in a dried state on the test strip and after pre-incubation with a sample. Pre-incubating the GNP conjugates and sample for 30 s was found to decrease the detection limit by 60-fold from 330 ng∙mL−1 to 5.4 ng∙mL−1 in comparison with conventional LFIA. The developed method was successfully tested for its ability to detect PVY in infected and uninfected potato leaves. The quantitative results of the proposed LFIA with pre-incubation were confirmed by ELISA, and resulted in a correlation coefficient of 0.891. The proposed approach is rapid, simple, and preserves the main advantages of LFIA as a non-laboratory diagnostic method.

scholarly journals Development and Application of a Novel Rapid and Throughput Method for Broad-Spectrum Anti-Foodborne Norovirus Antibody Testing

2021 ◽  
Vol 12 ◽  
Author(s):  
Yueting Zuo ◽  
Liang Xue ◽  
Junshan Gao ◽  
Yingyin Liao ◽  
Yueting Jiang ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Broad Spectrum ◽  
High Throughput Screening ◽  
Indirect Elisa ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Assay ◽  
Antibody Testing ◽  
Coefficients Of Variation ◽  
P Proteins

Foodbone norovirus (NoV) is the leading cause of acute gastroenteritis worldwide. Candidate vaccines are being developed, however, no licensed vaccines are currently available for managing NoV infections. Screening for stimulated antibodies with broad-spectrum binding activities can be performed for the development of NoV polyvalent vaccines. In this study, we aimed to develop an indirect enzyme-linked immunosorbent assay (ELISA) for testing the broad spectrum of anti-NoV antibodies. Capsid P proteins from 28 representative NoV strains (GI.1–GI.9 and GII.1–GII.22 except GII.11, GII.18, and GII.19) were selected, prepared, and used as coating antigens on one microplate. Combined with incubation and the horseradish peroxidase chromogenic reaction, the entire process for testing the spectrum of unknown antibodies required 2 h for completion. The intra-assay and inter-assay coefficients of variation were less than 10%. The new method was successfully performed with monoclonal antibodies and polyclonal antibodies induced by multiple antigens. In conclusion, the indirect ELISA assay developed in this study had a good performance of reliability, convenience, and high-throughput screening for broad-spectrum antibodies.

Stress-related compounds in xylem fluid of blight-diseased citrus containing Fusarium solani naphthazarin toxins and their effects on the host

1995 ◽  
Vol 41 (6) ◽  
pp. 515-524 ◽  
Author(s):  
S. Nemec
Keyword(s):  
Fusarium Solani ◽  
Protein Concentration ◽  
Phenylalanine Ammonia Lyase ◽  
Fold Increase ◽  
Enzyme Linked Immunosorbent Assay ◽  
Acid Oxidase ◽  
Tree Roots ◽  
Xylem Fluid ◽  
Stress Metabolites ◽  
Related Compounds

Naphthazarin toxins of Fusarium solani were detected and quantified by competitive enzyme-linked immunosorbent assay (ELISA) in xylem fluid of scaffold roots from blight-diseased trees. These toxins alter plant metabolic activity; this study examined their effects on xylem health by measuring physiological components in xylem fluid. Protein concentration in fluid was positively correlated with increases in toxin concentration. In fluid containing about 100 μg∙L−1 toxin, total amino acids reached levels 2.5 to 3.0 times greater than those in fluid containing no detectable toxin; asparagine, glutamic acid, proline, glycine, and arginine were the most abundant. Levels of phenylalanine ammonia-lyase, polyphenol oxidase, chlorogenic acid oxidase, and superoxide dismutase activity did not increase in xylem fluid containing toxin, which may be a reason why vascular discoloration did not occur. Xylem fluid containing about 20 μg∙L−1 toxin was associated with a 9-fold increase in total phenolics and a 15-fold increase in peroxidase. Peroxidases were predominantly anionic and may function in defense. Some of these peroxidases may function as lignases, releasing phenolic and other constituents from cells and cell walls. These toxins are known to enhance membrane permeability, which may be the main reason for the accumulation of these stress metabolites in xylem fluid. These data explain the disruption of hydraulic conductivity in blight tree roots and the eventual physiological breakdown of roots on diseased trees.Key words: phytotoxins, isomarticin, ELISA, fungi, roots.

Enzyme-Linked Immunosorbent Assay for Streptomycin in Animal Feeds

2013 ◽  
Vol 651 ◽  
pp. 280-283
Author(s):  
Hui Ying Zhang ◽  
Jun Ping Wang
Keyword(s):  
Detection Limit ◽  
Bovine Serum Albumin ◽  
Immunosorbent Assay ◽  
Polyclonal Antibody ◽  
Bovine Serum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Animal Feeds ◽  
Inhibition Concentration ◽  
Coefficients Of Variation ◽  
Bovine Serum Albumin Conjugate

Polyclonal antibody against streptomycin was prepared by using a streptomycin–bovine serum albumin conjugate for the immunization of rabbits. Using this antibody, we developed quantitative assays for streptomycin by means of an indirect competitive enzyme-linked immunosorbent assay (icELISA). Fifty percent inhibition concentration (IC50) for the antibody was 3.6 ng/ml. The detection limit was 0.4 ng/ml. The average of recoveries for all samples was 86.14% and the coefficients of variation of intra- and inter-assays were below 18%. The detection limit using the kit was 15 ng/ml in animal feeds.

Development of an immunological method for studying the incorporation of ripening enzymes in cheese curd

1996 ◽  
Vol 63 (1) ◽  
pp. 141-149 ◽  
Author(s):  
Edith Laloy ◽  
Jean-Christophe Vuillemard ◽  
Mouhsine El Abboudi ◽  
Ismael Fliss ◽  
Eric De Grace ◽  
...  
Keyword(s):  
Detection Limit ◽  
Immunosorbent Assay ◽  
Total Protein ◽  
Lactobacillus Casei ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein G ◽  
Immunological Method ◽  
Liposome Suspension ◽  
Cheese Curd

SummaryPolyclonal antibodies raised against partly purified aminopeptidase were specific to cell-free extracts ofLactobacillus caseisubsp.pseudoplantarumUL 137, and did not cross react with any other proteins in Cheddar cheese, at least during the first week of maturation. A sandwich enzyme-linked immunosorbent assay (ELISA) was developed in order to quantify cell-free extracts in the cheese curd, and was also used to determine the efficiency of encapsulation of cell-free extracts in liposomes. This method was very sensitive and exhibited a detection limit of ∼10 μg total protein/g cheese and ∼1 μg total protein/ml liposome suspension. The efficiency of encapsulation of cell-free extracts into liposomes was ∼55–60%. The retention of liposome-encapsulated cell-free extracts was ∼14 times that of non-encapsulated extracts.

Determination of Ochratoxin A in Feed by Immunoaffinity Cleanup and Liquid Chromatography

2013 ◽  
Vol 96 (3) ◽  
pp. 599-602 ◽  
Author(s):  
Ping Ding ◽  
Ziyou Mi ◽  
Yali Hou ◽  
Yigang He ◽  
Jianhua Xie
Keyword(s):  
Liquid Chromatography ◽  
Detection Limit ◽  
Ochratoxin A ◽  
Cyanogen Bromide ◽  
Polyclonal Antibodies ◽  
Immunoaffinity Column ◽  
Low Levels ◽  
Sepharose 4B ◽  
Feed Samples

Abstract A method using LC was developed for determination of ochratoxin A (OTA) in feeds. The extracted samples were cleaned up by an immunoaffinity column prepared by covalently coupling polyclonal antibodies against OTA to cyanogen bromide-activated Sepharose 4B. The eluates were determined by LC with fluorescence detection. Recoveries of OTA from fortified samples of 1–10 μg/kg levels ranged from 84.3 to 90.0%, with CVs of 3.3–7.8%. The detection limit was 0.045 μg/kg based on an S/N of 3:1. A total of 65 feed samples were screened for OTA with the proposed method. The results showed that only nine samples were contaminated with OTAs at low levels. The presented method was successfully applied to quantify OTAs in real feed samples.

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