Enzyme-linked immunosorbent assay for serum amyloid A apolipoprotein with use of specific antibodies against synthetic peptides.

1988 ◽  
Vol 34 (9) ◽  
pp. 1767-1771 ◽  
Author(s):  
R Saïle ◽  
C Delpierre ◽  
P Puchois ◽  
G Hocke ◽  
C Cachera ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Synthetic Peptides ◽  
Solid Phase ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Antibodies ◽  
Amyloid A ◽  
Microtiter Plates ◽  
Sensitivity Specificity ◽  
As Solid Phase

Abstract We describe a noncompetitive enzyme-linked immunosorbent assay for amyloid A apolipoprotein in human serum (apo SAA) in which specific antibodies against synthetic peptides are used. Microtiter plates were used as solid phase and coated with affinity-purified antibodies raised against SAA1-(95-104) peptide. After incubation with delipidated plasmas, the bound apo SAA was revealed by labeled antibodies raised against SAA1-(58-69) peptide. The assay offers several advantages over existing techniques: sensitivity, specificity, simplicity, and non-use of radioisotopes. Results correlate well with those by a nephelometric method in which polyclonal antibodies are used.

Enzyme-linked immunosorbent assay for detection of antibodies to extractable nuclear antigens in systemic lupus erythematosus, with nylon as solid phase.

1983 ◽  
Vol 29 (5) ◽  
pp. 823-827 ◽  
Author(s):  
M Ishaq ◽  
R Ali
Keyword(s):  
Systemic Lupus Erythematosus ◽  
Immunosorbent Assay ◽  
Lupus Erythematosus ◽  
Solid Phase ◽  
Enzyme Linked Immunosorbent Assay ◽  
Prolonged Storage ◽  
Antibody Activity ◽  
Systemic Lupus ◽  
Nuclear Antigens ◽  
As Solid Phase

Abstract In this enzyme-linked immunosorbent assay (ELISA) for detection of antibodies against extractable nuclear antigens (ENA) in sera of patients with systemic lupus erythematosus (SLE), nylon is used as solid phase for antigen binding instead of the commonly used polystyrene surface. Optimal conditions for activation of the nylon beads, antigen coating, and other relevant factors have been investigated. We compared the incidence of anti-ENA antibodies in SLE, using chromogenic and fluorogenic enzyme substrates. Of SLE patients, 54% were positive for anti-ENA antibodies when chromogenic substrate was used as compared with 68% for fluorogenic substrate. Antibody activity against Sm and RNP antigens was distinguished on the basis of ribonuclease sensitivity of the RNP antigen. The method described offers advantages such as decreased background activity, increased surface area, facility for prolonged storage of antigen-coated solid phase, and miniaturization of the assay.

scholarly journals Development of an enzyme-linked immunosorbent assay (ELISA) for the detection of specific antibodies against an H7N7 and an H3N8 equine influenza virus

1984 ◽  
Vol 93 (3) ◽  
pp. 609-620 ◽  
Author(s):  
M. S. Denyer ◽  
J. R. Crowther ◽  
R. C. Wardley ◽  
R. Burrows
Keyword(s):  
Influenza Virus ◽  
Immunosorbent Assay ◽  
Solid Phase ◽  
Allantoic Fluid ◽  
Influenza Viruses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Equine Influenza Virus ◽  
Serum Samples ◽  
Specific Antibodies ◽  
Equine Influenza

SummaryThis paper describes a solid-phase microtitre plate enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to equine influenza viruses. Using egg-grown influenza viruses as the antigens attached to the solid phase, crossreactions were observed between an H7N7 equine virus (designated A1) and an H3N8 equine influenza virus (designated A2) when untreated antisera were tested. Absorption of antisera with egg-grown A/Porcine/Shope/1/33 influenza virus eliminated cross-reactive antibodies so that specific detection of anti-equine influenza A1 or A2 antibodies was possible.Examination of horse sera following vaccination with A1 and/or A2 isolates showed that antibodies were produced against antigen associated with egg allantoic fluid as well as against virus. Such antibodies were eliminated following the absorption of antisera with porcine influenza virus. Results using sera from horses with known vaccination histories confirmed that the ELISA preferentially detected antibodies homologous to the antigen attached to the solid phase and methods to evaluate the current serological state of individual horses by relating the titres of specific antibodies against equine influenza A1 and A2 isolates are shown. This ELISA provides a simple and rapid method of assessing specific antibodies from horse sera and offers advantages over the ‘routine’ HI and SRH assessments since it gives high precision, is economical of reagents and has the capacity to handle large numbers of serum samples.

scholarly journals Production of Monoclonal Antibodies Specific for the i and 1,2 Flagellar Antigens of Salmonella typhimurium and Characterization of Their Respective Epitopes

1998 ◽  
Vol 64 (12) ◽  
pp. 5033-5038 ◽  
Author(s):  
N. de Vries ◽  
K. A. Zwaagstra ◽  
J. H. J. Huis in’t Veld ◽  
F. van Knapen ◽  
F. G. van Zijderveld ◽  
...  
Keyword(s):  
Amino Acids ◽  
Monoclonal Antibodies ◽  
Salmonella Typhimurium ◽  
Immunosorbent Assay ◽  
Specific Antibody ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Antibodies ◽  
Minimal Area

ABSTRACT Salmonella typhimurium expresses two antigenically distinct flagellins, each containing a different H antigen (i and 1,2), the combination of which is highly specific for this serotype. In this study, overlapping recombinant flagellin fragments were constructed from the fliC(H:i) and fljB (H:1,2) flagellin genes, and the expression products were tested for binding to H antigen-specific monoclonal and polyclonal antibodies. A minimal area, 86 amino acids for H:i and 102 amino acids for H:1,2, located in the central variable domain of each flagellin was required for the binding of serotype-specific antibodies, providing further evidence for the presence of a discontinuous H epitope. Two peptides comprising these areas were shown to be highly suitable for application as antigens in an enzyme-linked immunosorbent assay detecting S. typhimurium-specific antibody.

Polyaniline-dacron composite as solid phase in enzyme linked immunosorbent assay forYersinia pestis antibody detection

2001 ◽  
Vol 56 (2) ◽  
pp. 257-260 ◽  
Author(s):  
Raquel Andrade Lima Co�lho ◽  
Guilherme Marques Pimentel Santos ◽  
Paula Helena Santos Azev�do ◽  
Georgia de Assis Jaques ◽  
Walter Mendes Azevedo ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Solid Phase ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
As Solid Phase

scholarly journals Polyvinyl alcohol-glutaraldehyde as solid-phase in Elisa for Schistosomiasis

1997 ◽  
Vol 39 (3) ◽  
pp. 155-158 ◽  
Author(s):  
Aureci M. ARAÚjO ◽  
Gustavo H.T.S. BARBOSA ◽  
José Ricardo P. DINIZ ◽  
Elizabeth MALAGUEÑO ◽  
Walter M. AZEVEDO ◽  
...  
Keyword(s):  
Polyvinyl Alcohol ◽  
Schistosoma Mansoni ◽  
Immunosorbent Assay ◽  
Reliable Method ◽  
Solid Phase ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antigen Preparation ◽  
Conventional Elisa ◽  
Set Up ◽  
As Solid Phase

Soluble adult Schistosoma mansoni antigen preparation (SWAP) was covalently fixed onto polyvinyl alcohol-glutaraldehyde discs and an enzyme linked-immunosorbent assay (ELISA) was set up. The best conditions for the assay were established and it was found that small amount of antigen such as 1.5 µg was required. A comparison between this procedure and the conventional ELISA was proceeded. A reliable method of antigen immobilization was achieved and the low prices of the employed reagents are economically attractive

Detection of Bovine Milk in Ovine Milk by a Sandwich Enzyme-Linked Immunosorbent Assay (ELISA)

1991 ◽  
Vol 54 (5) ◽  
pp. 366-371 ◽  
Author(s):  
TERESA GARCÍA ◽  
ROSARIO MARTÍN ◽  
ELENA RODRÍGUEZ ◽  
JUAN I. AZCONA ◽  
BERNABÉ SANZ ◽  
...  
Keyword(s):  
Optical Density ◽  
Immunosorbent Assay ◽  
Polyclonal Antibodies ◽  
Sandwich Elisa ◽  
Whey Proteins ◽  
Bovine Milk ◽  
Microtiter Plate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Antibodies ◽  
Specific Antigens

A sandwich ELISA (enzyme-linked immunosorbent assay) was developed for the detection of defined amounts of bovine milk (1–30%) in ovine milk. Polyclonal antibodies were raised in rabbits against bovine whey proteins (BWP). Resultant antibodies were affinity purified by immunoadsorption of the crude antiserum onto columns containing immobilized ovine, caprine, and BWP, followed by elution of the bovine milk specific antibodies (anti-BWP) from the column containing the bovine proteins. The specific anti-BWP antibodies bound to the wells of a microtiter plate were used to capture the BWP from milk mixtures. Further immunorecognition of the captured proteins was attained with the same specific antibodies conjugated to biotin. ExtrAvidin-peroxidase was used to detect the biotinylated antibodies bound to their specific antigens. Subsequent enzymic conversion of substrate gave clear optical density differences when assaying mixtures containing variable amounts of bovine milk.

Indirect Enzyme-Linked Immunosorbent Assay for Detection of Aflatoxin B1 in Corn and Peanut Butter

1984 ◽  
Vol 47 (4) ◽  
pp. 263-266 ◽  
Author(s):  
TITAN S.L. FAN ◽  
FUNS. CHU
Keyword(s):  
Immunosorbent Assay ◽  
Aflatoxin B1 ◽  
Solid Phase ◽  
Peanut Butter ◽  
Enzyme Linked Immunosorbent Assay ◽  
Corn Meal ◽  
Additional Advantage ◽  
Microtiter Plates ◽  
Sample Extract ◽  
Direct Elisa

An indirect enzyme-linked immunosorbent assay (ELISA) for aflatoxin B1 (AFB1) was developed. The method involves coating AFB1-polylysine conjugate on microtiter plates as immobilized antigen, followed by incubation with free toxin standard or sample extract and anti-aflatoxin antibody from rabbits. The amount of antibody bound to the solid phase was determined by subsequent incubation with a secondary antibody conjugated with an enzyme, i.e., goat anti-rabbit IgG-horseradish peroxidase conjugate, and reaction with the chromogenic substrate. Aflatoxins were extracted from peanut butter and corn meal samples according to the BF and CB method of the Association of Official Analytical Chemists, respectively. Extracts without column cleanup treatment were dissolved in assay buffer for subsequent ELISA. Using this technique, 79.5 to 98.6% and 68 to 97% of AFB1 added in the range of 5 to 40 (μg/kg to the corn meal and peanut butter samples were recovered, respectively. The indirect ELISA achieved the same sensitivity and specificity for AFB1 as that obtained from the direct ELISA, with an additional advantage that much less antibody (100 times less) was required for the assay.

scholarly journals Dot enzyme-linked immunosorbent assay (dot-ELISA) for schistosomiasis diagnosis using dacron as solid-phase

1999 ◽  
Vol 32 (2) ◽  
pp. 139-143 ◽  
Author(s):  
Silvia Maria Lucena Montenegro ◽  
Joanne D'arc Bezerra da Silva ◽  
Maria Edileuza Felinto de Brito ◽  
Luiz Bezerra de Carvalho Junior
Keyword(s):  
Immunosorbent Assay ◽  
Solid Phase ◽  
Population Surveillance ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Individuals ◽  
Antigen Preparation ◽  
Endemic Areas ◽  
The Difference ◽  
Dot Elisa ◽  
As Solid Phase

Dacron and nitrocellulose were evaluated as matrices for the dot enzyme linked immunosorbent assay (dot-ELISA) for schistosomiasis and compared to indirect immunofluorescence (IMF). Titration of sera from 18 schistosomiasis patients against soluble worm antigen preparation (SWAP) was carried out and sera from healthy individuals from non-endemic areas were used as controls. The IMF was less sensitive than the dot-ELISAs, although the difference was not statistically significant (p > 0.05). The dot-ELISA based on nitrocellulose was as sensitive as that using dacron. Stability did not differ between nitrocellulose and dacron. Specificity was lower when dacron was used than when nitrocellulose was used, although the difference was not statistically significant (p > 0.05). In conclusion, this work showed that nitrocellulose and dacron performed similarly in dot-ELISA, suggesting that they may be used alternatively in population surveillance in endemic areas.

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