Comparison of monoclonal antibody performance characteristics for the detection of two representatives of A- and B-trichothecenes: T-2 toxin and deoxynivalenol

2010 ◽  
Vol 3 (3) ◽  
pp. 233-238 ◽  
Author(s):  
S. Baumgartner ◽  
M. Führer ◽  
R. Krska
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Animal Health ◽  
Type B ◽  
Fusarium Spp ◽  
Food And Feed ◽  
Type A Trichothecenes ◽  
Line Selection ◽  
Immunological Test ◽  
Cell Line Selection

Mycotoxins are secondary metabolites produced by fungi belonging mainly to the Aspergillus, Penicillium and Fusarium genera. They represent a relevant source of danger to human and animal health causing food- and feedborne intoxication. One group, produced by Fusarium spp., are the trichothecenes, of which T-2 toxin belongs to the type-A trichothecenes and deoxynivalenol to the type-B trichothecenes. As these mycotoxins are ubiquitous, the testing of products is required to keep our food and feed safe. For this purpose, sensitive and reliable tests are needed to detect contaminations. One detection possibility is an immunoanalytical based test which needs antibodies as reagents. Cell culture facilities allow cell line selection and production of monoclonal antibodies for further immunological test development. Especially for mycotoxins antibody development for further use in immunoassays is a crucial task. T-2 toxin and deoxynivalenol specific monoclonal antibodies were developed and further characterised to test stability and solvent resistance properties. Especially cross-reactivities were determined to related mycotoxins also belonging to the trichothecene family, e.g. HT-2 toxin or 3-acetyldeoxynivalenol.

scholarly journals Type A Trichothecene Diacetoxyscirpenol-Induced Emesis Corresponds to Secretion of Peptide YY and Serotonin in Mink

Toxins ◽  
2020 ◽  
Vol 12 (6) ◽  
pp. 419
Author(s):  
Qinghua Wu ◽  
Kamil Kuca ◽  
Eugenie Nepovimova ◽  
Wenda Wu
Keyword(s):  
Food Safety ◽  
Potential Risk ◽  
Peptide Yy ◽  
Animal Health ◽  
Type A ◽  
European Food Safety Authority ◽  
Cereal Grains ◽  
Blood Collection ◽  
Type B ◽  
Type A Trichothecenes

The trichothecene mycotoxins contaminate cereal grains and have been related to alimentary toxicosis resulted in emetic response. This family of mycotoxins comprises type A to D groups of toxic sesquiterpene chemicals. Diacetoxyscirpenol (DAS), one of the most toxic type A trichothecenes, is considered to be a potential risk for human and animal health by the European Food Safety Authority. Other type A trichothecenes, T-2 toxin and HT-2 toxin, as well as type B trichothecene deoxynivalenol (DON), have been previously demonstrated to induce emetic response in the mink, and this response has been associated with the plasma elevation of neurotransmitters peptide YY (PYY) and serotonin (5-hydroxytryptamine, 5-HT). However, it is found that not all the type A and type B trichothecenes have the capacity to induce PYY and 5-HT. It is necessary to identify the roles of these two emetogenic mediators on DAS-induced emesis. The goal of this study was to determine the emetic effect of DAS and relate this effect to PYY and 5-HT, using a mink bioassay. Briefly, minks were fasted one day before experiment and given DAS by intraperitoneally and orally dosing on the experiment day. Then, emetic episodes were calculated and blood collection was employed for PYY and 5-HT test. DAS elicited robust emetic responses that corresponded to upraised PYY and 5-HT. Blocking the neuropeptide Y2 receptor (NPY2R) diminished emesis induction by PYY and DAS. The serotonin 3 receptor (5-HT3R) inhibitor granisetron totally restrained the induction of emesis by serotonin and DAS. In conclusion, our findings demonstrate that PYY and 5-HT have critical roles in DAS-induced emetic response.

What about the 'other' Fusarium mycotoxins?

2009 ◽  
Vol 2 (2) ◽  
pp. 181-192 ◽  
Author(s):  
M. Jestoi ◽  
M. Kokkonen ◽  
S. Uhlig
Keyword(s):  
Current Knowledge ◽  
Fusarium Species ◽  
Animal Health ◽  
Natural Occurrence ◽  
Fusarium Mycotoxins ◽  
Fusarium Spp ◽  
Natural Toxins ◽  
Toxic Metabolites ◽  
Food And Feed ◽  
Very High

Most Fusarium species are capable of producing mycotoxins that may cause adverse effects on human or animal health. The most commonly studied Fusarium mycotoxins include trichothecenes, zearalenone and fumonisins. However, it seems that nearly all of the most prevalent Fusarium species infecting grains are also capable of producing other toxic metabolites. The existing studies, although exiguous, have clearly demonstrated that other toxic metabolites of Fusarium spp. are also present in our foods and feeds, occasionally at very high levels. It is apparent that since mycotoxins, including these 'other' metabolites, are natural toxins, they cannot be completely eliminated from food and feed chains. However, scientific studies are needed to determine their true significance. Thus, the mechanism and level of toxicity as well as presence and concentration levels will have to be fully clarified. In this paper, we briefly review the prevalence of the dominant Fusarium species contaminating maize and small-grain cereals worldwide, and the current knowledge on the biological activity as well as the natural occurrence of their selected less-known toxic metabolites. Additionally, the significance of these 'other' Fusarium mycotoxins is discussed.

scholarly journals PSII-22 Trends in Mycotoxin Contamination in United States Corn

2020 ◽  
Vol 98 (Supplement_3) ◽  
pp. 172-173
Author(s):  
Erika Hendel ◽  
Shelby Ramirez ◽  
Paige Gott ◽  
G Raj Murugesan ◽  
Ursula Hofstetter
Keyword(s):  
Animal Health ◽  
Corn Silage ◽  
Mitigation Strategies ◽  
Fungal Metabolites ◽  
Mycotoxin Contamination ◽  
Food And Feed ◽  
Type A Trichothecenes ◽  
Low Levels ◽  
Feed Safety ◽  
New Crop

Abstract Mycotoxins are harmful secondary fungal metabolites and are of key concern to food and feed safety globally. These toxins are detrimental to animal health and can compromise animal performance even at low levels. Classic signs such as decreased feed intake and vomiting used as indicators for exposure overlook other costs of mycotoxicosis, including increasing the frequency and severity of disease via immune suppression, inciting inflammation, and modulating the gastrointestinal environment. This survey examines initial samples of the 2019 crop with previous year trends. New crop corn samples were submitted from September 2019 and consisted of corn (46%), corn silage (50%), and corn byproduct (4%). Samples were analyzed utilizing the liquid chromatography and tandem mass spectrometry (LC-MS/MS) method for six major mycotoxin groups: aflatoxins (Afla), type A trichothecenes (A-Trich), type B trichothecenes (B-Trich), fumonisins (FUM), zearalenone (ZEN), and ochratoxin-A (OTA). Data are presented for major mycotoxin classes in Table 1. Fewer samples are available thus far compared to the fall of 2018 (50 samples in 2019 vs. 135 samples in 2018), thus risk profile of this crop year is likely to change as the sample pool expands. Co-occurrence (≥ 2 mycotoxins) has decreased compared to 2018. The prevalence of B-Trich decreased compared with previous years, but levels are similar to 2018. Prevalence and levels of ZEN decreased from 2018, and are similar to 2017, while FUM is similar in prevalence to 2018, but average ppb numerically increased. As of yet, no Alfa has been detected; however, corn stored with higher moisture content has increased the risk for storage toxins. Mycotoxin risk of this harvest season is still coming into focus as harvest delays have affected sample submission. Due to continued risk of multi-mycotoxin contamination, multiple mitigation strategies are needed beyond just adsorption, including biotransformation, support of the immune system and liver function.

scholarly journals Fusarium Phytotoxin Trichothecenes Have an Elicitor-Like Activity in Arabidopsis thaliana, but the Activity Differed Significantly Among Their Molecular Species

2006 ◽  
Vol 19 (5) ◽  
pp. 512-520 ◽  
Author(s):  
Takumi Nishiuchi ◽  
Daisuke Masuda ◽  
Hideo Nakashita ◽  
Kazuya Ichimura ◽  
Kazuo Shinozaki ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Signaling Pathway ◽  
Host Plants ◽  
Type A ◽  
Phytopathogenic Fungi ◽  
Type B ◽  
Fusarium Spp ◽  
Callose Deposition ◽  
Mitogen Activated Protein ◽  
Type A Trichothecenes

Phytopathogenic fungi such as Fusarium spp. synthesize trichothecene family phytotoxins. Although the type B trichothecene, deoxynivalenol (DON), is thought to be a virulence factor allowing infection of plants by their trichothecene-producing Fusarium spp., little is known about effects of trichothecenes on the defense response in host plants. Therefore, in this article, we investigated these effects of various trichothecenes in Fusarium-susceptible Arabidopsis thaliana. Necrotic lesions were observed in Arabidopsis leaves infiltrated by 1 μM type A trichothecenes such as T-2 toxin. Trichothecene-induced lesions exhibited dead cells, callose deposition, generation of hydrogen peroxide, and accumulation of salicylic acids. Moreover, infiltration by trichothecenes caused rapid and prolonged activation of two mitogen-activated protein kinases and induced expression of both PR-1 and PDF1.2 genes. Thus, type A trichothecenes trigger the cell death by activation of an elicitor-like signaling pathway in Arabidopsis. Although DON did not have such an activity even at 10 μM, translational inhibition by DON was observed at concentrations above 5 μM. These results suggested that DON is capable of inhibiting translation in Arabidopsis cells without induction of the elicitor-like signaling pathway.

scholarly journals 170 Trends in mycotoxin contamination in the united states corn

2019 ◽  
Vol 97 (Supplement_2) ◽  
pp. 93-94 ◽  
Author(s):  
Shelby Curry ◽  
Erika G Hendel ◽  
Paige Gott ◽  
G R Murugesan ◽  
Ursula Hofstetter-Schähs
Keyword(s):  
Animal Health ◽  
The United States ◽  
Fungal Species ◽  
Mitigation Strategies ◽  
Fungal Metabolites ◽  
Mycotoxin Contamination ◽  
Increased Risk ◽  
Food And Feed ◽  
Type A Trichothecenes ◽  
Feed Safety

Abstract Mycotoxins are harmful secondary fungal metabolites and are of key concern to food and feed safety globally. In addition to compromised performance, mycotoxins negatively impact animal health. Although classic signs such as decreased feed intake and vomiting are known in the field as indicators for exposure, mycotoxins act as predisposing factors for diseases by immune suppression, causing inflammation, and modulating the gastrointestinal environment, even at low levels. This survey presents mycotoxin levels of corn samples from the 2018 harvest and compares these levels with those in previous years. New crop corn samples from various sources, were submitted starting from mid-August 2018, and consisted of corn (70%), corn silage (18%), and corn byproduct (12%). Samples were analyzed utilizing the liquid chromatography and tandem mass spectrometry (LC-MS/MS) method for six major mycotoxin groups: aflatoxins (Afla), type A trichothecenes (A-Trich), type B trichothecenes (B-Trich), fumonisins (FUM), zearalenone (ZEN), and ochratoxin-A (OTA). Data are presented for major mycotoxin classes in Table 1. The majority of samples contained at least 1 detectable mycotoxin with co-occurrence (≥ 2 mycotoxins) similar to 2017, and less than 2016. Prevalence of B-Trich has decreased compared with previous years, but average ppb is similar to 2017. Prevalence and average ppb of ZEN are similar to 2017, while FUM has increased in both prevalence and average ppb. Alfa prevalence has increased and average ppb is numerically higher than the previous two years. The preliminary results from the 2018 corn harvest suggest a continued risk from mycotoxins produced by Fusarium fungal species, and a potential increased risk of Afla compared to previous years. Because of the risk of multi-mycotoxin contamination in corn samples thus far, multiple mitigation strategies are needed beyond just adsorption, including biotransformation support of the immune system and liver function.

Rapid quantification and in situ detection of nitrifying bacteria in biofilms by monoclonal antibody method

2000 ◽  
Vol 41 (4-5) ◽  
pp. 301-308 ◽  
Author(s):  
N. Noda ◽  
H. Ikuta ◽  
Y. Ebie ◽  
A. Hirata ◽  
S. Tsuneda ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Fluorescent Antibody ◽  
Nitrifying Bacteria ◽  
Fluorescent Antibody Technique ◽  
Antibody Technique ◽  
Antibody Method ◽  
In Situ Detection ◽  
Rapid Quantification

Fluorescent antibody technique by the monoclonal antibody method is very useful and helpful for the rapid quantification and in situ detection of the specific bacteria like nitrifiers in a mixed baxterial habitat such as a biofilm. In this study, twelve monoclonal antibodies against Nitrosomonas europaea (IFO14298) and sixteen against Nitrobacter winogradskyi (IFO14297) were raised from splenocytes of mice (BALB/c). It was found that these antibodies exhibited little cross reactivity against various kinds of heterotrophic bacteria. The direct cell count method using monoclonal antibodies could exactly detect and rapidly quantify N. europaea and N. winogradskyi. Moreover, the distribution of N. europaea and N. winogradskyi in a biofilm could be examined by in situ fluorescent antibody technique. It was shown that most of N. winogradskyi existed near the surface part and most of N. europaea existed at the inner part of the polyethylene glycol (PEG) gel pellet, which had entrapped activated sludge and used in a landfill leachate treatment reactor. It was suggested that this monoclonal antibody method was utilized for estimating and controlling the population of nitrifying bacteria as a quick and favorable tool.

Therapeutic Monoclonal Antibodies in Clinical Practice against Cancer

2020 ◽  
Vol 20 (16) ◽  
pp. 1895-1907
Author(s):  
Navgeet Kaur ◽  
Anju Goyal ◽  
Rakesh K. Sindhu
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Clinical Practice ◽  
Therapeutic Agent ◽  
Hematologic Malignancies ◽  
Immune Checkpoints ◽  
Malignant Cells ◽  
Main Target ◽  
Therapeutic Monoclonal Antibodies ◽  
Cancerous Cells

The importance of monoclonal antibodies in oncology has increased drastically following the discovery of Milstein and Kohler. Since the first approval of the monoclonal antibody, i.e. Rituximab in 1997 by the FDA, there was a decline in further applications but this number has significantly increased over the last three decades for various therapeutic applications due to the lesser side effects in comparison to the traditional chemotherapy methods. Presently, numerous monoclonal antibodies have been approved and many are in queue for approval as a strong therapeutic agent for treating hematologic malignancies and solid tumors. The main target checkpoints for the monoclonal antibodies against cancer cells include EGFR, VEGF, CD and tyrosine kinase which are overexpressed in malignant cells. Other immune checkpoints like CTLA-4, PD-1 and PD-1 receptors targeted by the recently developed antibodies increase the capability of the immune system in destroying the cancerous cells. Here, in this review, the mechanism of action, uses and target points of the approved mAbs against cancer have been summarized.

Evaluation of the Efficiency of Protein A Affinity Chromatography to Purify a Monoclonal Antibody for Cancer Treatment and its Purity Analysis

2020 ◽  
Vol 7 (2) ◽  
pp. 121-133
Author(s):  
Ayesha Akhtar ◽  
Shivakumar Arumugam ◽  
Shoaib Alam
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Affinity Chromatography ◽  
Protein A ◽  
Exchange Chromatography ◽  
Size Exclusion ◽  
High Yield ◽  
Staphylococcal Protein A ◽  
Purification Step ◽  
Purity Analysis

Background:: Protein A affinity chromatography is often employed as the most crucial purification step for monoclonal antibodies to achieve high yield with purity and throughput requirements. Introduction:: Protein A, also known as Staphylococcal protein A (SPA) is found in the cell wall of the bacteria staphylococcus aureus. It is one of the first discovered immunoglobulin binding molecules and has been extensively studied since the past few decades. The efficiency of Protein A affinity chromatography to purify a recombinant monoclonal antibody in a cell culture sample has been evaluated, which removes 99.0% of feed stream impurities. Materials and Method:: We have systematically evaluated the purification performance by using a battery of analytical methods SDS-PAGE (non-reduced and reduced sample), Cation Exchange Chromatography (CEX), Size-exclusion chromatography (SEC), and Reversed phased-Reduced Chromatography for a CHO-derived monoclonal antibody. Results and Discussion:: The analytical test was conducted to determine the impurity parameter, Host Cell Contaminating Proteins (HCP). It was evaluated to be 0.015ng/ml after the purification step; while initially, it was found to be 24.431ng/ml. Conclusion:: The tests showed a distinct decrease in the level of different impurities after the chromatography step. It can be concluded that Protein A chromatography is an efficient step in the purification of monoclonal antibodies.

scholarly journals Development of a New Highly Selective Monoclonal Antibody against Preferentially Expressed Antigen in Melanoma (PRAME) and Identification of the Target Epitope by Bio-Layer Interferometry

2021 ◽  
Vol 22 (6) ◽  
pp. 3166
Author(s):  
Jwala Priyadarsini Sivaccumar ◽  
Antonio Leonardi ◽  
Emanuela Iaccarino ◽  
Giusy Corvino ◽  
Luca Sanguigno ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Western Blotting ◽  
Cancer Biomarkers ◽  
Trypsin Digestion ◽  
Protein Fragments ◽  
Target Epitope ◽  
Potential Activity ◽  
Antibody Epitopes

Background: Monoclonal antibodies (mAbs) against cancer biomarkers are key reagents in diagnosis and therapy. One such relevant biomarker is a preferentially expressed antigen in melanoma (PRAME) that is selectively expressed in many tumors. Knowing mAb’s epitope is of utmost importance for understanding the potential activity and therapeutic prospective of the reagents. Methods: We generated a mAb against PRAME immunizing mice with PRAME fragment 161–415; the affinity of the antibody for the protein was evaluated by ELISA and SPR, and its ability to detect the protein in cells was probed by cytofluorimetry and Western blotting experiments. The antibody epitope was identified immobilizing the mAb on bio-layer interferometry (BLI) sensor chip, capturing protein fragments obtained following trypsin digestion and performing mass spectrometry analyses. Results: A mAb against PRAME with an affinity of 35 pM was obtained and characterized. Its epitope on PRAME was localized on residues 202–212, taking advantage of the low volumes and lack of fluidics underlying the BLI settings. Conclusions: The new anti-PRAME mAb recognizes the folded protein on the surface of cell membranes suggesting that the antibody’s epitope is well exposed. BLI sensor chips can be used to identify antibody epitopes.

A novel method for the non-chromatographic purification of technetium-99m-labeled monoclonal antibodies: a study with B72.3 monoclonal antibody

1994 ◽  
Vol 21 (2) ◽  
pp. 171-178 ◽  
Author(s):  
Howard S. Rosenzweig ◽  
Girish N. Ranadive ◽  
Troy Seskey ◽  
Michael W. Epperly ◽  
William D. Bloomer
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Chromatographic Purification ◽  
Technetium 99M ◽  
Novel Method
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