scholarly journals Analysis of DDT Isomers with Enzyme-Linked Immunosorbent Assay and Optical Immunosensor Based on Rat Monoclonal Antibodies as Biological Recognition Elements

2010 ◽  
Vol 93 (1) ◽  
pp. 44-58 ◽  
Author(s):  
Petra M Krämer ◽  
Cristina M Weber ◽  
Stephan Forster ◽  
Peter Rauch ◽  
Elisabeth Kremmer
Keyword(s):  
Monoclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Lambda Light Chain ◽  
Optical Readout ◽  
Biological Recognition ◽  
Optical Immunosensor ◽  
Recognition Elements ◽  
Field Excitation ◽  
Phosphate Buffered Saline ◽  
Surface Water Samples

Abstract New rat monoclonal antibodies (mAbs) for DDT [1,1,1-trichloro-2,2-bis (4-chlorophenyl) ethane], namely DDT 7C12, DDT 1C1, and DDT 1B2, were developed, characterized, and applied in ELISA both in coating antigen and in enzyme-tracer format. The latter used horseradish peroxidase (HRP) or glucose oxidase as enzymes. The lowest concentration of p,p-DDT was determined with mAb DDT 7C12 and DDT-hapten HRP, with a test midpoint (IC50) of 0.5 ± 0.2 µg/L (n = 10) in 40 mM PBS (phosphate-buffered saline). The mouse anti-rat immunoglobulin lambda-light chain mAb LA1B12 was used as capture mAb. The best IC50 for o,p´-DDT in 40 mM PBS was 1.0 ± 0.3 µg/L (n = 12) and was obtained with mAb DDT 1C1 and DDT-hapten HRP, whereas mAb DDT 1B2 was very selective for p,p-DDT with an IC50 of 4.2 ± 1.6 µg/L (in 40 mM PBS, n = 9). An optical immunosensor was optimized and applied for the analysis of DDT (or DDT equivalents). This immunosensor consists of a bench-top optical readout device and disposable sensor chips, which include the fluidic system. Evanescent field excitation and emission of the fluorophore Oyster<sup/>-645 was used. An IC50 for p,p´-DDT [in 5 (v/v) isopropanol in 40 mM PBS] of 4 µg/L was obtained using DDT 7C12-Oyster-645. ELISA and immunosensor were used for the analysis of p,p-DDT in unspiked and spiked surface water samples. Within the working ranges of these immunotechniques, recoveries ranged from 80 to 120.

scholarly journals Epitope Mapping of Monoclonal Antibodies to Calreticulin Reveals That Charged Amino Acids Are Essential for Antibody Binding

Antibodies ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 31
Author(s):  
Ann Christina Bergmann ◽  
Cecilie Kyllesbech ◽  
Rimantas Slibinskas ◽  
Evaldas Ciplys ◽  
Peter Højrup ◽  
...  
Keyword(s):  
Amino Acids ◽  
Monoclonal Antibodies ◽  
Epitope Mapping ◽  
Biological Function ◽  
Specific Binding ◽  
Enzyme Linked Immunosorbent Assay ◽  
Potential Therapeutic Target ◽  
Charged Amino Acids ◽  
Chaperone Protein ◽  
Terminal Amino

Calreticulin is a chaperone protein, which is associated with myeloproliferative diseases. In this study, we used resin-bound peptides to characterize two monoclonal antibodies (mAbs) directed to calreticulin, mAb FMC 75 and mAb 16, which both have significantly contributed to understanding the biological function of calreticulin. The antigenicity of the resin-bound peptides was determined by modified enzyme-linked immunosorbent assay. Specific binding was determined to an 8-mer epitope located in the N-terminal (amino acids 34–41) and to a 12-mer peptide located in the C-terminal (amino acids 362–373). Using truncated peptides, the epitopes were identified as TSRWIESK and DEEQRLKEEED for mAb FMC 75 and mAb 16, respectively, where, especially the charged amino acids, were found to have a central role for a stable binding. Further studies indicated that the epitope of mAb FMC 75 is assessable in the oligomeric structure of calreticulin, making this epitope a potential therapeutic target.

scholarly journals Advances in emergent biological recognition elements and bioelectronics for diagnosing COVID-19

2021 ◽  
Vol 4 (1) ◽  
pp. 231-247
Author(s):  
Praopim Limsakul ◽  
Krit Charupanit ◽  
Chochanon Moonla ◽  
Itthipon Jeerapan
Keyword(s):  
Biological Recognition ◽  
Recognition Elements

scholarly journals A Survey of G6 and G10 Serotypes of Group a Bovine Rotaviruses from Diarrheic Beef and Dairy Calves using Monoclonal Antibodies in ELISA

1994 ◽  
Vol 6 (2) ◽  
pp. 175-181 ◽  
Author(s):  
A. Lucchelli ◽  
S. Y. Kang ◽  
M. K. Jayasekera ◽  
A. V. Parwani ◽  
D. H. Zeman ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
South Dakota ◽  
Dairy Herd ◽  
Dairy Calves ◽  
Enzyme Linked Immunosorbent Assay ◽  
Positive Stool ◽  
Field Samples ◽  
Group A Rotaviruses ◽  
Beef Herds ◽  
Group A

Group A bovine rotaviruses (BRV) have been identified worldwide as a major cause of diarrhea in the young of many species, including humans. Group A rotaviruses are classified into serotypes on the basis of the outer capsid proteins, VP7 (G types) and VP4 (P types). To date, there are 14 G types of group A rotaviruses, with G1, G6, G8, and G10 described for BRV isolates. In this study, G6- and G lo-specific monoclonal antibodies (MAbs) were used in an enzyme-linked immunosorbent assay (ELISA) for the G typing of BRV-positive stool samples from diarrheic beef and dairy calves from South Dakota, Ohio, Michigan, Nebraska, and Washington, USA, and Ontario, Canada. ELISA plates were coated using a broadly reactive VP7 MAb (Common 60) or with G6- or G10-specific MAbs. BRV-positive fecal samples were diluted and added to duplicate wells, followed by the addition of polyclonal guinea pig anti-group A rotavirus serum as the secondary antibody. Several reference G6 and G10 BRV strains as well as other G types previously reported in cattle (G1, G2, G3, G8) and BRV-negative samples were included as G type specificity and negative controls. From a total of 308 field samples analyzed, 79% (244/308) tested positive by the broadly reactive VP7 MAb; of these, 54% (131/244) were G6 positive, 14% (35/244) were G10 positive, 4% (9/244) were both G6 and G10 positive, and 28% (69/244) were G6 and G10 negative. The negative samples may represent additional or undefined serotypes. The 89 samples from South Dakota were further subdivided into samples from beef ( n = 43) or dairy ( n = 46) herds. G6 was more prevalent in beef herd samples (67%) than in dairy herd samples (47.5%). In addition, dairy herds had higher percentages of G10-positive samples (17.5%) G6-G10 double positives (10%), and untypable samples (25%) than did beef herds, in which the prevalence of G10 positive samples was 5.5%, G6-G10 double positives was 5.5%, and untypable samples was 22%. Application of the serotype ELISA for the analysis of additional BRV samples will provide further epidemiologic data on the distribution of BRV serotypes in beef or dairy cattle, an important consideration for the development of improved BRV vaccines.

Production and use of monoclonal antibodies for identification of Bradyrhizobium japonicum strains

1988 ◽  
Vol 34 (1) ◽  
pp. 88-92 ◽  
Author(s):  
D. Velez ◽  
J. D. Macmillan ◽  
L. Miller
Keyword(s):  
Monoclonal Antibodies ◽  
Bradyrhizobium Japonicum ◽  
Chemical Nature ◽  
Periodate Oxidation ◽  
Enzyme Linked Immunosorbent Assay ◽  
Gram Negative Bacteria ◽  
Bacterial Cells ◽  
Gram Negative ◽  
Polyclonal Antisera ◽  
Somatic Antigens

Thirteen murine hybridomas capable of producing monoclonal antibodies to somatic antigens on Bradyrhizobium japonicum were developed and an indirect enzyme-linked immunosorbent assay was used to test reactivity of the antibodies against 20 strains of B. japonicum. Although polyclonal antisera from mice immunized with strains of B. japonicum reacted with bacterial cells of all 20 strains, individual monoclonals were more specific. Some antibodies reacted with as few as 2 and one with as many as 11 strains. On the basis of reactivity with the set of 13 monoclonal antibodies, the 20 strains of B. japonicum could be divided arbitrarily into five groups. Three of five monoclonal antibodies tested reacted with bacteroids taken directly from soybean nodules. One monoclonal bound to cells of five species of Rhizobium, but none of the 13 reacted with gram-negative bacteria representing six other genera. Treatment of cells with reagents and heat indicated the chemical nature of the antigens to five of the monoclonals. Antigen reactive with one antibody was destroyed by periodate oxidation indicating that it was a polysaccharide. Two antigens were probably proteins as they could be digested by trypsin and denatured by heat. Two others were inactivated by all three treatments suggesting they were glycoproteins.

scholarly journals Enzyme-Linked Immunosorbent Assay Using Monoclonal Antibodies for Direct Serotyping of Enteric Adenoviruses in Feces

1991 ◽  
Vol 65 (5) ◽  
pp. 552-558 ◽  
Author(s):  
Kenji TAKAGI ◽  
Yasutaka YAMASHITA ◽  
Hiroo INOUYE ◽  
Mitsuaki OHSETO ◽  
Hiroko KUWABARA ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Enteric Adenoviruses

scholarly journals Immunomagnetic Separation and Coagglutination of Vibrio parahaemolyticus with Anti-Flagellar Protein Monoclonal Antibody

2008 ◽  
Vol 15 (10) ◽  
pp. 1541-1546 ◽  
Author(s):  
S. Datta ◽  
M. E. Janes ◽  
J. G. Simonson
Keyword(s):  
Staphylococcus Aureus ◽  
Monoclonal Antibody ◽  
Cell Suspension ◽  
Vibrio Parahaemolyticus ◽  
Vibrio Vulnificus ◽  
Immunomagnetic Separation ◽  
Enzyme Linked Immunosorbent Assay ◽  
Selective Isolation ◽  
Flagellar Protein ◽  
Phosphate Buffered Saline

ABSTRACT Mice were immunized by injection of Vibrio parahaemolyticus ATCC 17802 polar flagellin in order to produce monoclonal antibodies (mAbs). mAbs were analyzed by anti-H enzyme-linked immunosorbent assay using V. parahaemolyticus polar flagellar cores. The mAb exhibiting the highest anti-H titer was coated onto Cowan I Staphylococcus aureus cells at a concentration of 75 μg/ml cell suspension and used for slide coagglutination. Of 41 isolates identified genetically as V. parahaemolyticus, 100% coagglutinated with the anti-H mAb within 30 s, and the mAb did not react with 30 isolates identified as Vibrio vulnificus. A strong coagglutination reaction with V. parahaemolyticus ATCC 17802 was still observed when the S. aureus cells were armed with as little as 15 μg of mAb/ml S. aureus cell suspension. At this concentration, the mAb cross-reacted with three other Vibrio species, suggesting that they share an identical H antigen or antigens. The anti-H mAb was then used to optimize an immunomagnetic separation protocol which exhibited from 35% to about 45% binding of 102 to 103 V. parahaemolyticus cells in phosphate-buffered saline. The mAb would be useful for the rapid and selective isolation, concentration, and detection of V. parahaemolyticus cells from environmental sources.

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

2006 ◽  
Vol 13 (3) ◽  
pp. 420-422 ◽  
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.

Rapid detection of human rotavirus strains in stools by single-sandwich enzyme-linked immunosorbent assay systems using monoclonal antibodies

1989 ◽  
Vol 24 (1-2) ◽  
pp. 43-56 ◽  
Author(s):  
Giuseppe Gerna ◽  
Antonella Sarasini ◽  
Angela Di Matteo ◽  
Maurizio Parea ◽  
Maria Torseilini ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Immunosorbent Assay ◽  
Rapid Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Rotavirus
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