Analysis of the Kanamycin in Raw Milk Using the Suspension Array

With the monoclonal antibody against kanamycin being prepared successfully, a bead-based indirect competitive fluorescent immunoassay was developed to detect kanamycin in milk. The fact that there was no significant cross-reaction with other aminoglycoside antibiotics implied that the monoclonal antibody was highly specific for kanamycin. The limit of detection (LOD) and the 50% inhibition concentration (IC50) in raw milk were 3.2 ng/mL and 52.5 ng/mL, respectively. Using the method developed in this study, the kanamycin concentrations were monitored in raw milk after the intramuscular administration of kanamycin in sick cows. Compared to the conventional enzyme-linked immunosorbent assay (ELISA), the method using the suspension array system was more sensitive. The results obtained in the present study showed a good correlation with that of the ELISA.

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Enzyme-linked immunosorbent assay for triclocarban in aquatic environments

Water Science & Technology ◽

10.2166/wst.2015.366 ◽

2015 ◽

Vol 72 (10) ◽

pp. 1682-1691 ◽

Cited By ~ 3

Author(s):

Kun Zeng ◽

Yanmin Zou ◽

Jianxia Liu ◽

Wei Wei ◽

Meng Zhang ◽

...

Keyword(s):

Monoclonal Antibody ◽

Immunosorbent Assay ◽

Limit Of Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Aquatic Environments ◽

Satisfactory Accuracy ◽

Good Linear ◽

High Affinity ◽

Inhibition Concentration ◽

Optimized Conditions

A sensitive, competitive indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of triclocarban (TCC) in waters and sediments. Haptens were synthesized by derivatizing the paraposition of a phenyl moiety of TCC. The synthesized hapten was then coupled to bovine thyroglobulin to be used as an immunogen, based on which, a high affinity monoclonal antibody 4D5 was produced with the hybridoma technique. Under the optimized conditions, using the monoclonal antibody, excellent performances of the assay were obtained: satisfactory sensitivity (IC50 (50% inhibition concentration) value, 0.43 ng/mL; limit of detection, 0.05 ng/mL); good linear range (0.05–10 ng/mL); and satisfactory accuracy (recoveries 70.7–107% in waters; 74.8–98.3% in sediments). Furthermore, TCC was found with the concentration ranging from not detected to 422.12 ng/L in waters and from 6.68 ng/g to 78.67 ng/g in sediments in Yunliang River, Ancient Canal and Hongqiao Port in Zhenjiang City. In conclusion ELISA could be applied for monitoring TCC in aquatic environments.

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

Planta Medica ◽

10.1055/a-0578-8689 ◽

2018 ◽

Vol 84 (14) ◽

pp. 1038-1044 ◽

Cited By ~ 3

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.

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

Foods ◽

10.3390/foods10102398 ◽

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.

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Sensitive Measurement of Thrombopoietin by a Monoclonal Antibody Based Sandwich Enzyme-linked Immunosorbent Assay

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657725 ◽

1997 ◽

Vol 78 (04) ◽

pp. 1262-1267 ◽

Cited By ~ 46

Author(s):

Claudia C Folman ◽

Albert E G K von dem Borne ◽

Irma H J A M Rensink ◽

Winald Gerritsen ◽

C Ellen van der Schoot ◽

...

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Immunosorbent Assay ◽

Platelet Transfusion ◽

Large Scale ◽

Limit Of Detection ◽

Cross Reactivity ◽

Enzyme Linked Immunosorbent Assay ◽

Reliable Tool

SummaryIn this report a sensitive enzyme-linked immunosorbent assay (ELISA) for the measurement of plasma thrombopoietin (Tpo) is described that is solely based on monoclonal antibodies (MoAbs).The assay has an intra and inter-assay variance of 5-7% and 7-13%, respectively. Native and recombinant human Tpo (rhTpo) were recognized equally well, no cross reactivity with other cytokines was found and rhTpo added to plasma and serum was completely recovered. With the ELISA, Tpo concentrations in EDTA-anticoagulated plasma of all controls (n = 193) could be determined, since the limit of detection (2 ± 0.8 A.U./ml, mean ± sd) was lower than the concentration found in controls (11 ± 8 A.U./ml, mean ± sd; 2.5th-97.5th percentile: 4-32 A.U./ml). Tpo levels in serum were on average 3.4 times higher than in plasma.We showed in vivo that Tpo is bound by platelets, as in thrombocytopenic patients (n = 5) a platelet transfusion immediately led to a drop in plasma Tpo level, whereas in patients receiving chemotherapy the induced thrombocytopenia was followed by a rise in plasma Tpo levels.In summary, these results indicate that this ELISA is a reliable tool for Tpo measurements and is applicable for large scale studies.

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Use of a Monoclonal Antibody and Two Enzyme-Linked Immunosorbent Assay Formats for Detection and Quantification of the Substitution of Caprine Milk for Ovine Milk

Journal of Food Protection ◽

10.4315/0362-028x-60.8.973 ◽

1997 ◽

Vol 60 (8) ◽

pp. 973-977 ◽

Cited By ~ 20

Author(s):

ANA I. HAZA ◽

PALOMA MORALES ◽

ROSARIO MARTÍIN ◽

TERESA GARCÍIA ◽

GONZALO ANGUITA ◽

...

Keyword(s):

Monoclonal Antibody ◽

Immunosorbent Assay ◽

Indirect Elisa ◽

Limit Of Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Goat’S Milk ◽

Elisa Format ◽

Ewe's Milk ◽

Goat's Milk ◽

Detection And Quantification

A stable hybridoma cell line (B2B) has been produced that secretes a monoclonal antibody (MAb) specific for goat's milk αS2-casein. The MAb B2B was used in two enzyme-linked immunosorbent assay (ELISA) formats for the detection and quantification of the presence of goat's milk in ewe's milk. In the indirect ELISA format the limit of detection was 0.5 to 15% (vol/vol) substitution of goat's milk for ewe's milk. Afterwards, a competitive indirect ELISA was successfully developed for the detection of 0.25 to 15% (vol/vol) of goat's milk in ewe's milk. This competitive indirect ELISA is a very sensitive assay; it can be performed in less than 5 h and is not influenced by the heat treatment of milk.

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Novel monoclonal antibody-based sensitive enzyme-linked immunosorbent assay and rapid immunochromatographic strip for detecting aflatoxin M1 in milk

Food Control ◽

10.1016/j.foodcont.2016.01.036 ◽

2016 ◽

Vol 66 ◽

pp. 1-7 ◽

Cited By ~ 19

Author(s):

Biing-Hui Liu ◽

Kuang–Chun Chu ◽

Feng-Yih Yu

Keyword(s):

Monoclonal Antibody ◽

Immunosorbent Assay ◽

Aflatoxin M1 ◽

Enzyme Linked Immunosorbent Assay ◽

Immunochromatographic Strip

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Unreliable Measurement of Basophil Maximum Leukotriene Release with the Bühlmann CAST 2000 Enzyme-Linked Immunosorbent Assay Kit

Clinical and Vaccine Immunology ◽

10.1128/cvi.13.3.420-422.2006 ◽

2006 ◽

Vol 13 (3) ◽

pp. 420-422 ◽

Cited By ~ 6

Author(s):

S. E. Burastero ◽

C. Paolucci ◽

D. Breda ◽

G. Monasterolo ◽

R. E. Rossi ◽

...

Keyword(s):

Monoclonal Antibody ◽

Monoclonal Antibodies ◽

Immunosorbent Assay ◽

False Positive ◽

Positive Control ◽

Enzyme Linked Immunosorbent Assay ◽

Positive Results

ABSTRACT The Bühlmann CAST 2000 enzyme-linked immunosorbent assay is a potentially useful assay for measuring sulfidoleukotrienes released in vitro by allergen-challenged basophils. However, we observed that the positive-control reagent yielded positive signals in cell-free systems. These false-positive results depended on using a mouse anti-FcεRI monoclonal antibody and were prevented by degranulation-inducing reagents other than mouse monoclonal antibodies.

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