Comparative Detection of Fumonisin by HPLC, ELISA, and Immunocytochemical Localization in Fusarium Cultures

1995 ◽  
Vol 58 (6) ◽  
pp. 666-672 ◽  
Author(s):  
MARIA VICTORIA TEJADA-SIMON ◽  
LILIAN T. MAROVATSANGA ◽  
JAMES J. PESTKA
Keyword(s):  
Monoclonal Antibody ◽  
Fusarium Moniliforme ◽  
High Performance ◽  
Time Course ◽  
Fumonisin B1 ◽  
Fusarium Proliferatum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Immunocytochemical Localization ◽  
Large Deposits

Fumonisins are a group of mycotoxins that are elaborated by Fusarium moniliforme and Fusarium proliferatum and that have recently been associated with animal and human disease. In this study, the time course of fumonisin B1 (FB1) production in corn was monitored in five Fusarium cultures using high-performance liquid chromatography (HPLC), enzyme-linked immunosorbent assay (ELISA), and in situ localization by an enzyme-linked immunocytochemical technique (ELICT). Using HPLC on culture extracts prepared with 50% (vol/vol) acetonitrile in water, FB1 was detectable at 3 days with maximal FB1 (ranging from 230 to 3,000 ppm) occurring between 14 and 28 days. Although there was a positive correlation between FB1 detected by HPLC and ELISA, the latter consistently yielded higher results than HPLC. Maximal FB1 “equivalents” detected by ELISA ranged from 12,000 to 35,000 ppm. Following fixation of Fusarium from cultures, ELICT revealed the presence of large deposits indicative of fumonisin or fumonisin-like cross-reacting compounds in mycelia, microconidia, and microconidia. Prior to fixation, these compounds were extractable in 50% (vol/vol) acetonitrile in water. ELICT results qualitatively correlated with HPLC and ELISA over the time course of the cultures. Taken together, the results suggest that (a) ELISA or ELICT could be used for qualitative screening of FB1-producing cultures, and (b) in addition to FB1, the monoclonal antibody-based ELISA detected one or more compounds that structurally resemble FB1 and occur concurrently with FB1.

scholarly journals A comparison between enzyme immunoassay and HPLC for ochratoxin A detection in green, roasted and instant coffee

2007 ◽  
Vol 50 (2) ◽  
pp. 349-359 ◽  
Author(s):  
Simone Fujii ◽  
Elisabete Yurie Sataque Ono ◽  
Ricardo Marcelo Reche Ribeiro ◽  
Fernanda Garcia Algarte Assunção ◽  
Cássia Reika Takabayashi ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Ochratoxin A ◽  
High Performance ◽  
Screening Tool ◽  
Correlation Coefficients ◽  
Enzyme Linked Immunosorbent Assay ◽  
Elisa Assay ◽  
Quality And Safety ◽  
Green Coffee ◽  
Instant Coffee

An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for ochratoxin A (OTA) detection in green, roasted and instant coffees was developed using anti-OTA monoclonal antibody. Immunological reagents prepared were OTA-BSA (4.76 mg/mL), anti-OTA.7 MAb (2x10³-fold dilution) and HRP-anti IgG (10³-fold dilution). The detection limit was 3.73 ng OTA/g and correlation coefficients (r) between this immunoassay and high performance liquid chromatography were 0.98 for green coffee, 0.98 for roasted and 0.86 for instant. OTA levels detected by ic-ELISA were higher than by HPLC, with ELISA/HPLC ratio of 0.66 - 1.46 (green coffee), 0.96 - 1.11 (roasted) and 0.93 - 1.82 (instant). ELISA recoveries for OTA added to coffee (5 - 70 ng/g) were 81.53 % for green coffee, 46.73 % for roasted and 64.35 % for instant, while recoveries by HPLC were 80.54 %, 45.91 % and 55.15 %, respectively. Matrices interferences were minimized by samples dilution before carrying out the ELISA assay. The results indicate that MAb-based ic-ELISA could be a simple, sensitive and specific screening tool for OTA detection, contributing to quality and safety of coffee products.

Fumonisin Production in Corn by Toxigenic Strains of Fusarium moniliforme and Fusarium proliferatum

1994 ◽  
Vol 57 (6) ◽  
pp. 514-521 ◽  
Author(s):  
CHARLES W. BACON ◽  
PAUL E. NELSON
Keyword(s):  
Fusarium Moniliforme ◽  
Fruits And Vegetables ◽  
Animal Feed ◽  
Fusaric Acid ◽  
Fumonisin B1 ◽  
Fusarium Proliferatum ◽  
Food Crops ◽  
Ear Rot ◽  
Corn Products ◽  
Toxigenic Strains

The fungi Fusarium moniliforme Sheldon and Fusarium proliferatum (Matsushima) Nirenberg produce a series of toxins on corn which include the fumonisins of which fumonisin B1 and B2 are considered to have cancer promoting activity. Both fungi produce similar ratios of the fumonisins B1 to B2. Other mycotoxins produced include moniliformin, fusarin C and fusaric acid. Fumonisin B1 has been shown to be responsible for most of the toxicological affects observed from ingesting corn infected by toxigenic isolates of these fungi. The distribution of the two fungi is generally similar, although F. proliferatum is isolated more frequently from sorghum than corn. They occur worldwide on other food crops, such as rice, sorghum, millet, several fruits and vegetables. Both fungi are ear rot pathogens of corn, thus, mycotoxin production occurs under field conditions, although it also may occur in storage. One or both fungi may have a frequency of occurrence of 90% or higher in corn; 90% of the F. moniliforme isolates produce the fumonisins. On corn and corn products the range of concentrations reported is 0.3 to 330 μg/g of corn-based product. These concentrations include both corn-based animal feed and human foods.

A Monoclonal Antibody-Based Enzyme-Linked Immunosorbent Assay for Determination of Homoharringtonine

Planta Medica ◽  
2018 ◽  
Vol 84 (14) ◽  
pp. 1038-1044 ◽  
Author(s):  
Benyakan Pongkitwitoon ◽  
Seiichi Sakamoto ◽  
Rika Nagamitsu ◽  
Waraporn Putalun ◽  
Hiroyuki Tanaka ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Myeloid Leukemia ◽  
High Performance ◽  
Sodium Periodate ◽  
Limit Of Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Study Support ◽  
Mediated Oxidation

AbstractHomoharringtonine (HHT), also known as omacetaxine, is a natural compound found in the genus Cephalotaxus and is a promising pharmaceutical drug used for the treatment of chronic or accelerated phase chronic myeloid leukemia. As a tool for the quantitative determination of HHT, a specific monoclonal antibody against HHT (MAb 6A1) was generated by conjugates prepared via sodium periodate-mediated oxidation. The developed indirect competitive enzyme-linked immunosorbent assay (icELISA) using MAb 6A1 was found to be highly specific and sensitive with a limit of detection for HHT of 48.8 ng/mL. Validation assays to evaluate precision and accuracy of the method were conducted by the use of intra- and inter-assay analysis, recovery test, and comparison analysis between the amounts of HHT determined by ELISA and high-performance liquid chromatography. These results revealed that the established icELISA using MAb 6A1 is specific, sensitive, and reliable enough to be applied to the qualitative analysis for HHT. Furthermore, the results of this study support the usefulness of sodium periodate as a reagent for the conjugation between Cephalotaxus alkaloids and proteins for producing specific antibodies.

Fumonisin B1 Production and Growth of Fusarium moniliforme and Fusarium proliferatum on Maize, Wheat, and Barley Grain

1999 ◽  
Vol 64 (5) ◽  
pp. 921-924 ◽  
Author(s):  
S. Marin ◽  
N. Magan ◽  
J. Serra ◽  
A.J. Ramos ◽  
R. Canela ◽  
...  
Keyword(s):  
Fusarium Moniliforme ◽  
Fumonisin B1 ◽  
Barley Grain ◽  
Fusarium Proliferatum

Stability of Fumonisins in Thermally Processed Corn Products†

1998 ◽  
Vol 61 (8) ◽  
pp. 1030-1033 ◽  
Author(s):  
MAURICIO M. CASTELO ◽  
SUSAN S. SUMNER ◽  
LLOYD B. BULLERMAN
Keyword(s):  
High Performance ◽  
Average Rate ◽  
Fumonisin B1 ◽  
High Performance Liquid Chromatographic ◽  
Hplc Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Corn Products ◽  
Dry Heating ◽  
The Stability ◽  
Liquid Chromatographic

Little is known about the stability of fumonisins in corn-based foods during heating. This study investigated the effects of canning, baking, and roasting (dry heating) processes on the stability of fumonisins in artificially contaminated and naturally contaminated corn-based foods. All samples were analyzed for fumonisin levels by both a commercial enzyme-linked immunosorbent assay (ELISA) and a high-performance liquid chromatographic (HPLC) method. Canned whole-kernel corn showed a significant (P ≤ 0.05) decrease in fumonisins by both ELISA (15%) and HPLC (11%) analyses. Canned cream-style com and baked corn bread showed significant (P ≤ 0.05) decreases in fumonisin levels at an average rate of 9% and 48%, respectively, as analyzed by ELISA. Corn-muffin mix artificially contaminated with 5 μg of fumonisin B1 (FB1,) per g and naturally contaminated corn-muffin mix showed no significant (P ≤ 0.05) losses of fumonisins upon baking. Roasting cornmeal samples artificially contaminated with 5 μg of FB1, per g and naturally contaminated commeal samples at 218°C for 15 min resulted in almost complete loss of fumonisins.

Fumonisin B1 Production by Fusarium moniliforme and Fusarium proliferatum as Affected by Cycling Temperatures†

1999 ◽  
Vol 62 (12) ◽  
pp. 1456-1460 ◽  
Author(s):  
DOJIN RYU ◽  
CELESTIN MUNIMBAZI ◽  
LLOYD B. BULLERMAN
Keyword(s):  
Low Temperature ◽  
Temperature Stress ◽  
Constant Temperature ◽  
Fusarium Moniliforme ◽  
Fusarium Species ◽  
Fumonisin B1 ◽  
Fusarium Proliferatum ◽  
Low Temperature Stress ◽  
Maximum Production ◽  
Cycling Temperature

The effects of temperatures cycling between 5 and 20°C, 10 and 25°C, and 15 and 30°C on the production of fumonisin B1 (FB1) and ergosterol by Fusarium moniliforme and Fusarium proliferatum on rice was studied. Temperatures were cycled at 12-h intervals by manually moving cultures from one temperature to another. Constant temperature incubation at 25°C and a low temperature stress were compared with the cycling temperature incubations. Low temperature stress was achieved by incubating rice cultures at 25°C for 2 weeks followed by 15°C for 4 weeks. The maximum yields of FB1 were found to be 247 μg/g by F. moniliforme at temperatures that cycled between 10 and 25°C after 2 weeks and 284 μg/g by F. proliferatum when the temperatures cycled between 5 and 20°C after 6 weeks. Ergosterol content of the rice cultures was also monitored. Overall, the two Fusarium species showed differences in production of FB1 and ergosterol under the various temperature treatments. The most notable differences were that the temperature treatments that stimulated greatest FB1 production were different for each species: cycling temperatures between 10 and 25°C for F. moniliforme and cycling temperatures between 5 and 25°C for F. proliferatum. At most temperatures, F proliferatum produced more ergosterol than F. moniliforme. Maximum production of ergosterol by F. proliferatum occurred at 6 weeks, with temperatures that cycled between 10 and 25°C, whereas F. moniliforme produced maximum amounts of ergosterol at 6 weeks, with temperatures that cycled between 15 and 30°C.

scholarly journals Generation of monoclonal antibodies detecting specific epitopes in locust antennae

1993 ◽  
Vol 175 (1) ◽  
pp. 45-57
Author(s):  
J. Strotmann ◽  
I. Boekhoff ◽  
S. Goggerle ◽  
H. Breer
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibody ◽  
Protein Synthesis ◽  
Monoclonal Antibodies ◽  
Embryonic Development ◽  
High Molecular Weight ◽  
Time Course ◽  
Antigen Expression

1. Following a tissue-specific screening paradigm, monoclonal antibodies have been generated that interact with distinct subpopulations of cells in locust antennae. 2. Antigens were identified as high molecular weight components. 3. Immunoreactivity was not detectable during embryonic development, but rapidly appeared within a few hours of hatching. 4. The time course of antigen expression in antennal cells could be followed in situ as well as in vitro. 5. Expression of monoclonal antibody B14/6D2-like immunoreactivity was prevented by blocking protein synthesis with cycloheximide.

scholarly journals Preparation of Anti-Aristolochic Acid I Monoclonal Antibody and Development of Chemiluminescent Immunoassay and Carbon Dot-Based Fluoroimmunoassay for Sensitive Detection of Aristolochic Acid I

Foods ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2647
Author(s):  
Ai-Fen Ou ◽  
Zi-Jian Chen ◽  
Yi-Feng Zhang ◽  
Qi-Yi He ◽  
Zhen-Lin Xu ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
High Performance ◽  
Aristolochic Acid ◽  
Limit Of Detection ◽  
Inner Filter Effect ◽  
Enzyme Linked Immunosorbent Assay ◽  
Triple Quadrupole Mass Spectrometer ◽  
Assay Sensitivity ◽  
Chemiluminescent Immunoassay ◽  
Aristolochic Acid I

Aristolochic acid (AA) toxicity has been shown in humans regarding carcinogenesis, nephrotoxicity, and mutagenicity. Monitoring the AA content in drug homologous and healthy foods is necessary for the health of humans. In this study, a monoclonal antibody (mAb) with high sensitivity for aristolochic acid I (AA-I) was prepared. Based on the obtained mAb, a chemiluminescent immunoassay (CLEIA) against AA-I was developed, which showed the 50% decrease in the RLUmax (IC50) value of 1.8 ng/mL and the limit of detection (LOD) of 0.4 ng/mL. Carbon dots with red emission at 620 nm, namely rCDs, were synthesized and employed in conventional indirect competitive enzyme-linked immunosorbent assay (icELISA) to improve the assay sensitivity of a fluoroimmunoassay (FIA). Oxidized 3,3′′,5,5′′-tetramethylbenzidine dihydrochloride (oxTMB) can quench the emission of the rCDs through the inner-filter effect; therefore, the fluorescence intensity of rCDs can be regulated by the concentration of mAb. As a result, the assay sensitivity of FIA was improved by five-fold compared to CLEIA. A good relationship between the results of the proposed assays and the standard ultra-high performance liquid chromatography-triple quadrupole mass spectrometer (UPLC-QQQ-MS/MS) of real samples indicated good accuracy and practicability of CLEIA and FIA.

scholarly journals LM6-M: a high avidity rat monoclonal antibody to pectic α-1,5-L-arabinan

2017 ◽  
Author(s):  
Valérie Cornuault ◽  
Fanny Buffetto ◽  
Susan E Marcus ◽  
Marie-Jeanne Crépeau ◽  
Fabienne Guillon ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
High Sensitivity ◽  
In Vitro Assays ◽  
Enzyme Linked Immunosorbent Assay ◽  
High Avidity ◽  
Plant Cell Walls ◽  
Rhamnogalacturonan I ◽  
Phosphate Buffered Saline

Abstract1,5-arabinan is an abundant structural feature of side chains of pectic rhamnogalacturonan-I which is a matrix constituent of plant cell walls. The study of arabinan in cells and tissues is driven by putative roles for this polysaccharide in the generation of cell wall and organ mechanical properties. The biological function(s) of arabinan is still uncertain and high quality molecular tools are required to detect its occurrence and monitor its dynamics. Here we report a new rat monoclonal antibody, LM6-M, similar in specificity to the published rat monoclonal antibody LM6 (Willats et al. (1998) Carbohydrate Research 308: 149-152). LM6-M is of the IgM immunoglobulin class and has a higher avidity for α-1-5-L-arabinan than LM6. LM6-M displays high sensitivity in its detection of arabinan in in-vitro assays such as ELISA and epitope detection chromatography and in in-situ analyses.AbbreviationsAraArabinoseBSABovine Serum AlbuminGalGalactoseGalAGalacturonic acidEDCEpitope detection chromatographyELISAEnzyme-Linked Immunosorbent AssaymAbMonoclonal antibodyPBSPhosphate-buffered salineRhaRhamnoseRG-IRhamnogalacturonan-I

Incidence and Levels of Fusarium moniliforme, Fusarium proliferatum and Fumonisins in Corn and Corn-Based Foods and Feeds1

1994 ◽  
Vol 57 (6) ◽  
pp. 541-546 ◽  
Author(s):  
LLOYD B. BULLERMAN ◽  
WEI-YUN J. TSAI
Keyword(s):  
Fusarium Moniliforme ◽  
Fumonisin B1 ◽  
The United States ◽  
Fusarium Proliferatum ◽  
Corn Meal ◽  
Closely Related Species ◽  
Dent Corn ◽  
Fumonisin B2 ◽  
Low Levels ◽  
Corn Flakes

Fusarium moniliforme Sheldon occurs worldwide on corn intended for human and animal consumption. A closely related species Fusarium proliferatum also occurs frequently on corn. Yellow dent corn, white dent corn, white and yellow popcorn and sweetcorn may be contaminated. Both organisms are capable of producing a group of toxins known as fumonisins, of which Fumonisin B1 (FB1), Fumonisin B2 (FB2) and Fumonisin B3 (FB3) are most common. Fumonisins have been found in corn and corn-based foods worldwide. Fumonisins may be found in sound whole kernel corn at levels at or below 1.0 μg/g. By contrast animal disease problems begin to occur at fumonisin levels above 5.0 to 10.0 μg/g. Corn-based food products that have the most frequent and highest fumonisin levels, besides whole kernels, are corn meal and corn grits. In the United States, corn meal has been found contaminated with Fumonisin B1 at levels from 0.5 to 2.05 μg/g, and grits from 0.14 to 0.27 μg/g. Corn flakes, corn pops, corn chips and tortilla chips have been found negative for fumonisins. Popcorn, sweetcorn and hominy corn have been found contaminated with sporadic, low levels (0.01 to 0.08 μg/g) of fumonisins. The effects of processing on fumonisins in corn are still largely unknown. Heating may cause a loss of fumonisins in corn, but it may be a loss of detectability rather than degradation.

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