Development of an enzyme-linked immunosorbent assay of apolipoprotein E-AII complex in plasma

1991 ◽  
Vol 37 (9) ◽  
pp. 1645-1648 ◽  
Author(s):  
M Tozuka ◽  
Y Yoshida ◽  
J Tanigami ◽  
M Miyachi ◽  
T Katsuyama ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Lipoprotein Metabolism ◽  
Enzyme Linked Immunosorbent Assay ◽  
Simple Method ◽  
Coefficients Of Variation ◽  
Apo E ◽  
Rabbit Igg ◽  
Reference Serum ◽  
Capture Antibody

Abstract Measurement of apolipoprotein (apo) E-AII complex in human plasma is important in determining the role of apoE in lipoprotein metabolism. In this paper, we demonstrate a new and simple method to determine apoE-All complex by using an enzyme-linked immunosorbent assay. Anti-apoE IgG (goat) was used as a capture antibody, and captured apoE-All complexes were detected by an anti-apoAll (rabbit) horseradish peroxidase-conjugated anti-rabbit IgG (goat) system. With this method, apoE-All complex was specifically determined without the interference of apoAll and was not affected by apoE monomer less than 250 mg/L. The content of the complex in reference serum, a normolipidemic serum pooled from five subjects with phenotype E3/E3, was arbitrarily defined as 100%. The coefficients of variation were 3.5%-6.3% within assay and 8.8%-11.6% between assays.

scholarly journals Measurement of plasminogen activator inhibitor 1 in biologic fluids with a murine monoclonal antibody-based enzyme-linked immunosorbent assay

Blood ◽  
1988 ◽  
Vol 71 (1) ◽  
pp. 220-225 ◽  
Author(s):  
PJ Declerck ◽  
MC Alessi ◽  
M Verstreken ◽  
EK Kruithof ◽  
I Juhan-Vague ◽  
...  
Keyword(s):  
Plasminogen Activator ◽  
Immunosorbent Assay ◽  
Healthy Subjects ◽  
Plasminogen Activator Inhibitor ◽  
Enzyme Linked Immunosorbent Assay ◽  
Plasminogen Activator Inhibitor 1 ◽  
Platelet Poor Plasma ◽  
Activator Inhibitor ◽  
Pai 1

An enzyme-linked immunosorbent assay for plasminogen activator inhibitor-1 (PAI-1) in biologic fluids was developed on the basis of two murine monoclonal antibodies raised against PAI-1 purified from HT- 1080 fibrosarcoma cells. The lower limit of sensitivity of the assay in plasma is 2 ng/mL. The assay is 12 times less sensitive toward the PAI- 1/human tissue-type plasminogen activator (t-PA) complex as compared with free PAI-1. The intraassay, interassay, and interdilution coefficients of variation are 5.2%, 8.0%, and 7.1%, respectively. The level of PAI-1 in platelet-poor plasma of healthy subjects is 18 +/- 10 ng/mL (mean +/- SD, n = 45). In platelet-rich plasma after freezing and thawing, 92% of PAI-1 antigen is released from platelets, whereas only 8% is found in the corresponding platelet-poor plasma. In platelet-poor plasma from healthy subjects, a linear correlation (r = 0.80) was found between PAI activity and PAI-1 antigen. In plasma approximately two thirds of the PAI-1 antigen was functionally active, whereas only 5% of the PAI-1 antigen released from platelets was active. During pregnancy a progressive increase of PAI-1 antigen levels up to three- to sixfold the control value was observed. In plasma of patients with recurrent deep vein thrombosis, PAI-1 levels were 44 +/- 20 ng/mL (mean +/- SD, n = 7), during a clinically silent phase. Four of these patients had a level above 38 ng/mL (mean +/- 2 SD of normal). The present assay, based on stable and reproducible reagents, allows the specific determination of PAI-1 antigen in biologic fluids. It may facilitate interlaboratory comparisons and be useful for further investigations of the role of PAI-1 in clinical conditions associated with impaired fibrinolysis and/or a tendency to thrombosis and investigations of the role of PAI-1 in platelets.

scholarly journals Assignment of Additional Anticapsular Antibody Concentrations to the Neisseria meningitidis Group A, C, Y, and W-135 Meningococcal Standard Reference Serum CDC1992

2002 ◽  
Vol 9 (3) ◽  
pp. 725-726 ◽  
Author(s):  
Cheryl M. Elie ◽  
Patricia K. Holder ◽  
Sandra Romero-Steiner ◽  
George M. Carlone
Keyword(s):  
Quality Control ◽  
Disease Control ◽  
Neisseria Meningitidis ◽  
Immunosorbent Assay ◽  
Immunoglobulin G ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control And Prevention ◽  
Group A ◽  
Reference Serum ◽  
Centers For Disease Control

ABSTRACT We assigned additional enzyme-linked immunosorbent assay antibody concentrations (immunoglobulin G [IgG], IgM, and IgA, and total) to the Neisseria meningitidis standard reference serum CDC1992 for groups Y and W-135 to 12 Centers for Disease Control and Prevention quality control sera. These assignments will supplement previous assignments and will aid in the evaluation of present and developing vaccines.

scholarly journals Measurement of plasminogen activator inhibitor 1 in biologic fluids with a murine monoclonal antibody-based enzyme-linked immunosorbent assay

Blood ◽  
1988 ◽  
Vol 71 (1) ◽  
pp. 220-225 ◽  
Author(s):  
PJ Declerck ◽  
MC Alessi ◽  
M Verstreken ◽  
EK Kruithof ◽  
I Juhan-Vague ◽  
...  
Keyword(s):  
Plasminogen Activator ◽  
Immunosorbent Assay ◽  
Healthy Subjects ◽  
Plasminogen Activator Inhibitor ◽  
Enzyme Linked Immunosorbent Assay ◽  
Plasminogen Activator Inhibitor 1 ◽  
Platelet Poor Plasma ◽  
Activator Inhibitor ◽  
Pai 1

Abstract An enzyme-linked immunosorbent assay for plasminogen activator inhibitor-1 (PAI-1) in biologic fluids was developed on the basis of two murine monoclonal antibodies raised against PAI-1 purified from HT- 1080 fibrosarcoma cells. The lower limit of sensitivity of the assay in plasma is 2 ng/mL. The assay is 12 times less sensitive toward the PAI- 1/human tissue-type plasminogen activator (t-PA) complex as compared with free PAI-1. The intraassay, interassay, and interdilution coefficients of variation are 5.2%, 8.0%, and 7.1%, respectively. The level of PAI-1 in platelet-poor plasma of healthy subjects is 18 +/- 10 ng/mL (mean +/- SD, n = 45). In platelet-rich plasma after freezing and thawing, 92% of PAI-1 antigen is released from platelets, whereas only 8% is found in the corresponding platelet-poor plasma. In platelet-poor plasma from healthy subjects, a linear correlation (r = 0.80) was found between PAI activity and PAI-1 antigen. In plasma approximately two thirds of the PAI-1 antigen was functionally active, whereas only 5% of the PAI-1 antigen released from platelets was active. During pregnancy a progressive increase of PAI-1 antigen levels up to three- to sixfold the control value was observed. In plasma of patients with recurrent deep vein thrombosis, PAI-1 levels were 44 +/- 20 ng/mL (mean +/- SD, n = 7), during a clinically silent phase. Four of these patients had a level above 38 ng/mL (mean +/- 2 SD of normal). The present assay, based on stable and reproducible reagents, allows the specific determination of PAI-1 antigen in biologic fluids. It may facilitate interlaboratory comparisons and be useful for further investigations of the role of PAI-1 in clinical conditions associated with impaired fibrinolysis and/or a tendency to thrombosis and investigations of the role of PAI-1 in platelets.

scholarly journals Distribution of Xylella fastidiosa in Oaks in Florida and Its Association with Growth Decline in Quercus laevis

Plant Disease ◽  
1998 ◽  
Vol 82 (5) ◽  
pp. 569-572 ◽  
Author(s):  
E. L. Barnard ◽  
E. C. Ash ◽  
D. L. Hopkins ◽  
R. J. McGovern
Keyword(s):  
Immunosorbent Assay ◽  
Xylella Fastidiosa ◽  
Shoot Length ◽  
Enzyme Linked Immunosorbent Assay ◽  
Oak Decline ◽  
Widespread Occurrence ◽  
Leaf Scorch ◽  
Growth Decline ◽  
Quercus Laevis

A survey of more than 200 trees has documented the widespread occurrence of Xylella fastidiosa in Florida oak populations. The pathogen was detected readily via enzyme-linked immunosorbent assay in oaks exhibiting decline or leaf scorch symptoms and was infrequently detected in asymptomatic trees. It was also associated with reduced growth in Quercus laevis as measured by current-year shoot length. The occurrence of X. fastidiosa in Q. laevis and the evidence for its occurrence in Q. incana represent first reports for these oak hosts. The role of X. fastidiosa in oak decline scenarios deserves further attention.

scholarly journals Comparison of four functionalization methods of gold nanoparticles for enhancing the enzyme-linked immunosorbent assay (ELISA)

2017 ◽  
Vol 8 ◽  
pp. 244-253 ◽  
Author(s):  
Paula Ciaurriz ◽  
Fátima Fernández ◽  
Edurne Tellechea ◽  
Jose F Moran ◽  
Aaron C Asensio
Keyword(s):  
Gold Nanoparticles ◽  
Detection Limit ◽  
Immunosorbent Assay ◽  
Wheat Gluten ◽  
Diagnostic Technique ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rabbit Igg ◽  
Enzyme Molecules ◽  
Main Components ◽  
Limit Sensitivity

The enzyme-linked immunosorbent assay (ELISA) technique is based on the specific recognition ability of the molecular structure of an antigen (epitope) by an antibody and is likely the most important diagnostic technique used today in bioscience. With this methodology, it is possible to diagnose illness, allergies, alimentary fraud, and even to detect small molecules such as toxins, pesticides, heavy metals, etc. For this reason, any procedures that improve the detection limit, sensitivity or reduce the analysis time could have an important impact in several fields. In this respect, many methods have been developed for improving the technique, ranging from fluorescence substrates to methods for increasing the number of enzyme molecules involved in the detection such as the biotin–streptavidin method. In this context, nanotechnology has offered a significant number of proposed solutions, mainly based on the functionalization of nanoparticles from gold to carbon which could be used as antibody carriers as well as reporter enzymes like peroxidase. However, few works have focused on the study of best practices for nanoparticle functionalization for ELISA enhancement. In this work, we use 20 nm gold nanoparticles (AuNPs) as a vehicle for secondary antibodies and peroxidase (HRP). The design of experiments technique (DOE) and four different methods for biomolecule loading were compared using a rabbit IgG/goat anti-rabbit IgG ELISA model (adsorption, directional, covalent and a combination thereof). As a result, AuNP probes prepared by direct adsorption were the most effective method. AuNPs probes were then used to detect gliadin, one of the main components of wheat gluten, the protein composite that causes celiac disease. With this optimized approach, our data showed a sensitivity increase of at least five times and a lower detection limit with respect to a standard ELISA of at least three times. Additionally, the assay time was remarkably decreased.

The role of enzyme-linked immunosorbent assay D-dimer in patients with acute coronary syndrome presenting with normal cardiac enzymes

2006 ◽  
Vol 17 (8) ◽  
pp. 621-624 ◽  
Author(s):  
Ariella Bar-Gil Shitrit ◽  
Dan Tzivony ◽  
Yuval Shilon ◽  
Bernard Rudensky ◽  
Jacklin Sulkes ◽  
...  
Keyword(s):  
Acute Coronary Syndrome ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cardiac Enzymes ◽  
Coronary Syndrome ◽  
D Dimer

scholarly journals Detection of Antibodies against Epidermodysplasia Verruciformis-Associated Canine Papillomavirus 3 in Sera of Dogs from Europe and Africa by Enzyme-Linked Immunosorbent Assay

2008 ◽  
Vol 16 (1) ◽  
pp. 66-72 ◽  
Author(s):  
C. E. Lange ◽  
K. Tobler ◽  
C. Favrot ◽  
M. Müller ◽  
J. O. Nöthling ◽  
...  
Keyword(s):  
South Africa ◽  
Fusion Protein ◽  
Immunosorbent Assay ◽  
Clinical Manifestations ◽  
Epidermodysplasia Verruciformis ◽  
Capsid Proteins ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Antigens ◽  
Specific Antibodies

ABSTRACT The role of papillomaviruses (PVs) in the development of canine cancers is controversial. However, recently a novel canine PV (CPV3) was detected in a dog affected with a condition reminiscent of epidermodysplasia verruciformis (EV). The aim of the present study was to investigate the seroprevalence of CPV3 by using generic enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies against either canine oral PV (COPV) or CPV3. Therefore, the capsid proteins of both PV types were expressed as glutathione S-transferase fusion protein antigens and adsorbed to glutathione-casein-coated ELISA plates. After showing that PV type-specific antibodies could be detected in the sera from dogs with confirmed COPV or CPV3 infection, CPV3- and COPV-seropositive samples were detected in two sets of canine sera collected in Switzerland and South Africa, respectively. We found specific antibodies against COPV and CPV3 among the tested sera and also a large number that were positive for both antigens. The seroprevalences of PV antibodies of 21.9% (COPV) and 26.9% (CPV3) among the tested dogs from South Africa were higher than those among the dogs from Switzerland at 10.5% (COPV) and 1.3% (CPV3). Our data suggest a need for further CPV-related seroepidemiological surveys in different countries, especially in the context of clinical manifestations and possible breed predispositions. For this purpose, the newly developed ELISAs can be a useful tool.

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