Tween 20 soluble proteins of Mycoplasma hyopneumoniae as antigen for an enzyme linked immunosorbent assay

1980 ◽  
Vol 29 (3) ◽  
pp. 305-309 ◽  
Author(s):  
J. Nicolet ◽  
P. Paroz ◽  
S. Bruggmann
Keyword(s):  
Immunosorbent Assay ◽  
Mycoplasma Hyopneumoniae ◽  
Soluble Proteins ◽  
Tween 20 ◽  
Enzyme Linked Immunosorbent Assay

Sandwich ELISA for Detection of Pig Meat in Raw Beef Using Antisera to Muscle Soluble Proteins

1988 ◽  
Vol 51 (10) ◽  
pp. 790-798 ◽  
Author(s):  
ROSARIO MARTÍN ◽  
JUAN I. AZCONA ◽  
CARMEN CASAS ◽  
PABLO E. HERNÁNDEZ ◽  
BERNABÉ SANZ
Keyword(s):  
Horseradish Peroxidase ◽  
Immunosorbent Assay ◽  
Peroxidase Activity ◽  
Sandwich Elisa ◽  
Soluble Proteins ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Antibodies ◽  
Double Antibody ◽  
Colored Product ◽  
Pig Meat

A double-antibody sandwich ELISA (enzyme-linked immunosorbent assay) has been successfully developed for the detection of defined amounts of pig meat (1–50%) in raw beef. Antibodies against pig sarcoplasmic extracts were produced in rabbits. Pig-specific antibodies were affinity purified by removing antibodies which crossreacted with horse, chicken or beef extracts followed by immunoadsorption and elution from a pig-extract column. The ELISA involved capturing antigens in sarcoplasmic extracts with pig specific antibodies immobilized on 96-well plates, detecting bound antigen with pig specific, horseradish peroxidase-labeling antibody, and measuring peroxidase activity by the conversion of a clear substrate to a colored product.

Enzyme-linked Immunosorbent Assay for a Soluble Antigen of Renibacterium salmoninarum, the Causative Agent of Salmonid Bacterial Kidney Disease

1987 ◽  
Vol 44 (1) ◽  
pp. 183-191 ◽  
Author(s):  
R. J. Pascho ◽  
D. Mulcahy
Keyword(s):  
Immunosorbent Assay ◽  
Incubation Temperature ◽  
Tween 80 ◽  
Broth Culture ◽  
Tween 20 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Soluble Antigen ◽  
Renibacterium Salmoninarum ◽  
Bacterial Kidney Disease ◽  
Wash Buffer

A double-antibody enzyme-linked immunosorbent assay (ELISA) for detection of a soluble fraction of Renibacterium salmoninarum was developed from components extracted from the supernatant of an R. salmoninarum broth culture. The Costar® Serocluster™ EIA microplate gave the highest absorbance and signal-to-noise ratios among seven types tested. Including Tween 80 in the wash buffer resulted in higher absorbances than Tween 20 when antigen was present. Background absorbance did not increase when Tween 80 was added to the wash buffer, but did when Tween 80 replaced Tween 20 in antigen and conjugate diluents. Adsorption of coating antibody peaked within 4 h at 37 °C and 16 h at 4 °C. Antigen attachment to antibody-coated microplate wells depended more on incubation temperature than duration; we adopted a 3-h incubation at 25 °C. Conjugate incubation for longer than 1 h at 37 °C or 3 h at 25 °C resulted in unacceptable background levels. No cross-reactions resulted from heat-extracted antigens of 10 other species of bacteria. The optimized ELISA is a 6-h test that enables detection of levels of soluble antigen as low as 2–20 ng.

Detection of Melamine Using Commercial Enzyme-Linked Immunosorbent Assay Technology

2008 ◽  
Vol 71 (3) ◽  
pp. 590-594 ◽  
Author(s):  
ERIC A. E. GARBER
Keyword(s):  
Standard Deviation ◽  
Immunosorbent Assay ◽  
Limit Of Detection ◽  
Elisa Test ◽  
Tween 20 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Screening Methods ◽  
Commercial Enzyme ◽  
Dog Food ◽  
Test Kit

Recent cases of adulteration with melamine have led to the need for rapid and reliable screening methods. To meet this need, commercial enzyme-linked immunosorbent assay (ELISA) test kits for the detection of triazines were evaluated. The recently released Melamine Plate kit (Abraxis, Warminster, Pa.) displayed a limit of detection of 9 ng/ml for melamine in phosphate-buffered saline (PBS) and approximately 1 μg/ml for melamine added to dog food. An atrazine ELISA test kit produced by Abraxis required 0.2 mg/ml to generate a response more than four times the standard deviation from background. In contrast, with the EnviroGard Triazine Plate kit (Strategic Diagnostics, Inc., Newark, Del.), 1.5 mg/ml melamine in PBS generated a signal only one standard deviation from background, which was insufficient to define a limit of detection. Extraction based on dilution with 105 mM sodium phosphate/75 mM NaCl/2.5% nonfat milk/0.05% Tween 20 (UD) enabled detection of fivefold less melamine in dog food than did use of the procedure recommended by the manufacturer, which entailed extraction into 60% methanol, sonication, centrifugation, filtration, and further dilution into 10% methanol/PBS. Using the Abraxis Melamine ELISA, both extraction protocols yielded identical results with a dog food sample adulterated with mela-mine. The recovery of melamine spiked into gravy from dog food using UD was 74% ± 4%. In conclusion, the recently released Abraxis ELISA for melamine proved to be a useful alternative to more cumbersome methods.

Enzyme-Linked Immunosorbent Assay of Ochratoxin A in Wheat

1984 ◽  
Vol 67 (1) ◽  
pp. 45-49 ◽  
Author(s):  
Sungsoo C Lee ◽  
Fun S Chu
Keyword(s):  
Horseradish Peroxidase ◽  
Ochratoxin A ◽  
Immunosorbent Assay ◽  
Sodium Phosphate ◽  
Methanolic Extract ◽  
Tween 20 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Final Solution ◽  
Reverse Phase ◽  
Methanol Content

Abstract An enzyme-linked immunosorbent assay (ELISA) was developed for the quantitation of ochratoxin A amended to wheat. Ochratoxin A conjugated to horseradish peroxidase (HRP) was used as an enzyme marker in the assay. At toxin levels below 30 ppb, a cleanup treatment was necessary for ELISA. Among 3 cleanup methods tested (solvent partition, Sep-Pak cartridge treatment, solvent partition and cartridge treatment), reverse phase cartridge treatment was the most simple and effective. In the analysis, ochratoxin A was extracted from wheat with methanol. The methanolic extract was diluted with water to a final 10–15% methanol content, and then passed through a cartridge. Ochratoxin A was eluted from the cartridge with 85% methanol which was then concentrated. The final solution, in 0.1M, pH 7.5 sodium phosphate–Tween 20 buffer and 5% methanol, was then subjected to ELISA. ELISA allowed minimal detection of the toxin in wheat at the 1–2 ppb level after cleanup. Recoveries of toxin added to wheat samples in the 1.0–30 ppb range were 85–90% with standard deviations of 10–15%.

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