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.

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.

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.

Specific 3H radioimmunoassay with a monoclonal antibody for monitoring cyclosporine in blood.

1988 ◽  
Vol 34 (2) ◽  
pp. 257-260 ◽  
Author(s):  
P E Ball ◽  
H Munzer ◽  
H P Keller ◽  
E Abisch ◽  
J Rosenthaler
Keyword(s):  
Monoclonal Antibody ◽  
Bone Marrow ◽  
High Performance Liquid Chromatography ◽  
High Performance ◽  
Limit Of Detection ◽  
Parent Drug ◽  
Sample Volume ◽  
Specific Radioimmunoassay ◽  
The Mean ◽  
Blood Specimens

Abstract A specific radioimmunoassay involving a mouse monoclonal antibody to cyclosporine has been developed for monitoring the parent drug in blood. Pretreatment with methanol removes cyclosporine from the erythrocytes. The limit of detection is about 12 micrograms/L, sample volume is 50 microL of blood, and within- and between-assay CVs are less than 7%. Assay results correlated well with those obtained by "high-performance" liquid chromatography (HPLC) for liver (n = 42), for heart (n = 64), for bone-marrow (n = 36), and for kidney (n = 140). For blood specimens obtained from patients treated with cyclosporine postoperatively for as long as 65 months, the mean RIA/HPLC ratio in all with transplant indications was close to 1. Therefore, the specific radioimmunoassay apparently can be used instead of HPLC to measure the parent drug in blood.

scholarly journals QuEChERS pretreatment combined with high-performance liquid chromatography-tandem mass spectrometry for determination of aristolochic acids I and II in Chinese herbal patent medicines

RSC Advances ◽  
2020 ◽  
Vol 10 (42) ◽  
pp. 25319-25324
Author(s):  
Jinghe Zhang ◽  
Yinan Wang ◽  
Jing Sun ◽  
Guowei Zhou ◽  
Xiaojie Jiang ◽  
...  
Keyword(s):  
High Performance ◽  
Aristolochic Acid ◽  
Patent Medicines ◽  
Chinese Herbal ◽  
Aristolochic Acids ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Aristolochic Acid I ◽  
Carcinogenic Compound

Aristolochic acid I and II (AA I and II), a kind of nephrotoxic and carcinogenic compound, are widely added in Chinese herbal patent medicines though they have been banned due to their toxicity.

scholarly journals Change of Amino Acid Residues in Idiotypic Nanobodies Enhanced the Sensitivity of Competitive Enzyme Immunoassay for Mycotoxin Ochratoxin A in Cereals

Toxins ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 273
Author(s):  
Caixia Zhang ◽  
Weiqi Zhang ◽  
Xiaoqian Tang ◽  
Qi Zhang ◽  
Wen Zhang ◽  
...  
Keyword(s):  
Amino Acid ◽  
Ochratoxin A ◽  
Limit Of Detection ◽  
Basic Knowledge ◽  
Affinity Constant ◽  
Enzyme Linked Immunosorbent Assay ◽  
Amino Acid Residues ◽  
Assay Sensitivity ◽  
Coating Antigen ◽  
Relative Standard

Anti-idiotypic nanobodies, usually expressed by gene engineering protocol, has been shown as a nontoxic coating antigen for toxic compound immunoassays. We here focused on how to increase immunoassay sensitivity by changing the nanobody’s primary sequence. In the experiments, two anti-idiotype nanobodies against monoclonal antibody 1H2, which is specific to ochratoxin A, were obtained and named as nontoxic coating antigen 1 (NCA1) and nontoxic coating antigen 2 (NCA2). Three differences between the nanobodies were discovered. First, there are six amino acid residues (AAR) of changes in the complementarity determining region (CDR), which compose the antigen-binding site. One of them locates in CDR1 (I–L), two of them in CDR2 (G–D, E–K), and three of them in CDR3 (Y–H, Y–W). Second, the affinity constant of NCA1 was tested as 1.20 × 108 L mol−1, which is about 4 times lower than that of NCA2 (5.36 × 108 L mol−1). Third, the sensitivity (50% inhibition concentration) of NCA1 for OTA was shown as 0.052 ng mL−1, which was 3.5 times lower than that of nontoxic coating antigen 2 (0.015 ng mL−1). The results indicate that the AAR changes in CDR of the anti-idiotypic nanobodies, from nonpolar to polar, increasing the affinity constant may enhance the immunoassay sensitivity. In addition, by using the nontoxic coating antigen 2 to substitute the routine synthetic toxic antigen, we established an eco-friendly and green enzyme-linked immunosorbent assay (ELISA) method for rapid detection of ochratoxin A in cereals. The half-maximal inhibitory concentration (IC50) of optimized ELISA was 0.017 ng mL−1 with a limit of detection (LOD) of 0.003 ng mL−1. The optimized immunoassay showed that the average recoveries of spiked corn, rice, and wheat were between 80% and 114.8%, with the relative standard deviation (RSD) ranging from 3.1–12.3%. Therefore, we provided not only basic knowledge on how to improve the structure of anti-idiotype nanobody for increasing assay sensitivity, but also an available eco-friendly ELISA for ochratoxin A in cereals.

Evaluation of Performance of a Commercial Monoclonal Antibody–Based Fenitrothion Immunoassay and Application to Residual Analysis in Fruit Samples

2006 ◽  
Vol 69 (1) ◽  
pp. 191-198 ◽  
Author(s):  
EIKI WATANABE ◽  
KOJI BABA ◽  
HEESOO EUN ◽  
TOMOHITO ARAO ◽  
YASUO ISHII ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Dynamic Range ◽  
Limit Of Detection ◽  
Close Correlation ◽  
Gas Chromatography Mass Spectrometry ◽  
Enzyme Linked Immunosorbent Assay ◽  
High Recovery ◽  
Elisa Kit ◽  
Fruit Samples ◽  
Standard Solutions

The performance of a commercially available enzyme-linked immunosorbent assay (ELISA) kit containing highly sensitive monoclonal antibodies against fenitrothion was assessed. The experimentally estimated dynamic range (0.087 to 2 ng/g) agreed with that established by the kit manufacturer (0.075 to 1 ng/g). The linearity of the standard curve produced for the kit-assembled standard solutions (slope = −0.3829) agreed with that of the curve produced for the self-made standard solutions (slope =−0.3619). The sensitivity (I50 value) and the limit of detection for the kit were 0.23 and 0.033 ng/g, respectively. Selectivity of the ELISA indicates that the monoclonal antibody can readily distinguish the target pesticide from other structurally related analogs and some metabolites (oxon forms), with the exception of ethyl O-(p-nitrophenyl) phenyl phosphonothionate (EPN), parathion-methyl, and parathion. Methanol was the best organic solvent for the kit, with optimal sensitivity observed at a final concentration in the well of 10% (vol/vol) or less. Matrix interferences were minimized by direct dilution with water (60-fold) of the methanolic extracts from apple and peach samples. To extract fenitrothion from these two agricultural products as simply and rapidly as possible, three extraction methods were used. The extraction method that involved shaking by hand for 3 min was the best among the three methods. High recovery percentages (116.6% for apple and 110.8% for peach) were obtained. Validation of the ELISA method was carried out using the gas chromatography–mass spectrometry method, resulting in high recovery and close correlation of results (r > 0.95). These findings strongly suggest that the ELISA kit may be routinely employed for on-site fenitrothion screening of fruit samples.

Development of an Enzyme-Linked Immunosorbent Assay for Determination Of Miroestrol Using an Anti-miroestrol Monoclonal Antibody

Planta Medica ◽  
2017 ◽  
Vol 83 (10) ◽  
pp. 855-861 ◽  
Author(s):  
Tharita Kitisripanya ◽  
Supaluk Krittanai ◽  
Orapin Udomsin ◽  
Kamonthip Jutathis ◽  
Jukrapun Komaikul ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Estrogenic Activity ◽  
Limit Of Detection ◽  
Simple Procedure ◽  
Enzyme Linked Immunosorbent Assay ◽  
Efficacy And Safety ◽  
Limit Of Quantitation ◽  
Estrogenic Action ◽  
White Kwao Krua

AbstractMiroestrol is a chromene with potent estrogenic activity present in Pueraria candollei, commonly known as White Kwao Krua. Although this compound is only present in low amounts in the plant, it plays an important role in the estrogenic action of P. candollei products. As a tool for further studies about the efficacy and safety of P. candollei as a phytoestrogenic supplement, we generated a novel monoclonal antibody against miroestrol. This anti-miroestrol monoclonal antibody was used to develop an immunoassay for the determination of miroestrol content, which can be used for quality control purposes of P. candollei. The developed ELISA against miroestrol has a calibration range of 10–780 ng/mL miroestrol, a limit of detection of 3.5 ng/mL, and a limit of quantitation of 12.2 ng/mL. According to the validation analysis, the established ELISA is precise, accurate, specific, and sensitive for miroestrol detection in plants. Furthermore, the anti-miroestrol monoclonal antibody was used to prepare an immunoaffinity column for the isolation of miroestrol from the tuberous root of P. candollei. The column provides a simple procedure for miroestrol isolation, with a capacity of 3.91 µg of miroestrol per 1 mL of immunogel.

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.

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.

Capture immunoassay for the diagnosis of bovine mastitis using a monoclonal antibody to polymorphonuclear granulocytes

1992 ◽  
Vol 59 (2) ◽  
pp. 123-133 ◽  
Author(s):  
Catherine A. O'Sullivan ◽  
Patrick J. Joyce ◽  
Teresa Sloan ◽  
Alan G. Shattock
Keyword(s):  
Monoclonal Antibody ◽  
Somatic Cell ◽  
Bovine Milk ◽  
Bovine Mastitis ◽  
Cell Counting ◽  
Enzyme Linked Immunosorbent Assay ◽  
Assay Sensitivity ◽  
Milk Samples ◽  
Cell Counts ◽  
Direct Elisa

SummaryA direct capture enzyme-linked immunosorbent assay (ELISA) was developed to measure elevated polymorphonuclear granulocyte (PMN) antigens using horseradish peroxidase (EC 1.11.1.7) conjugated rabbit polyclonal anti-PMN antisera and a monoclonal antibody specific for PMN cells. Optical densities obtained in the ELISA were used to predict the cell counts of milk samples. Predicted counts were not significantly different from actual somatic cell counts (SCC). In a total of 156 bovine milk samples the correlation coefficient between somatic cell counting, taking > 500000 cells/ml as being indicative of mastitis, and the assay was 0·94, yielding an assay sensitivity of 95·2% and a specificity of 97·3%. In further trials the ELISA could detect elevated PMN antigens in milk with SCC as low as 100000 cells/ml. The results indicate that the monoclonal antibody based direct ELISA has excellent potential in the detection and determination of bovine mastitis.

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