Differences between changes of antigen-specific IgE antibody level and those of IgG antibody level estimated by enzyme-linked immunosorbent assay(ELISA) in rats following sensitization with DNP-Ascaris.

SUMMARYEnzyme-linked immunosorbent assay (ELISA) tests were used to measure IgG antibody levels in 2638 New Zealand children who had been immunized with the triple vaccine DTP. The percentage of children immune to diphtheria decreased with age. The percentage of children immune to tetanus varied from 67.1 to 55.0%. The percentage of children with measurable antibody to pertussis increased with age. The mean percentages of children with measurable antibody or immunity to one or more DTP components were 34.2% (with 3 components), 34.4% (2 components), and 78.1% (1 component). It appears the immunization strategy for diphtheria and tetanus is satisfactory for herd immunity in New Zealand children. However, the current pertussis strategy may not be providing adequate immunity to 5-year-olds in this country.

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Amplification of the enzyme-linked immunosorbent assay for measuring allergen-specific IgE and IgG antibody

Clinical & Experimental Allergy ◽

10.1111/j.1365-2222.1981.tb02170.x ◽

1981 ◽

Vol 11 (6) ◽

pp. 523-531 ◽

Cited By ~ 20

Author(s):

W. J. METZGER ◽

J. E. BUTLER ◽

PRISCILLA SWANSON ◽

E. REINDERS* ◽

H. B. RICHERSON

Keyword(s):

Immunosorbent Assay ◽

Specific Ige ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibody

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Development and Validation of 13-plex Luminex-Based Assay for Measuring Human Serum Antibodies to Streptococcus pneumoniae Capsular Polysaccharides

mSphere ◽

10.1128/msphere.00128-18 ◽

2018 ◽

Vol 3 (4) ◽

pp. e00128-18 ◽

Cited By ~ 9

Author(s):

Danka Pavliakova ◽

Peter C. Giardina ◽

Soraya Moghazeh ◽

Shite Sebastian ◽

Maya Koster ◽

...

Keyword(s):

Human Serum ◽

Lower Limit ◽

Immunosorbent Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibodies ◽

Igg Antibody ◽

Correlate Of Protection ◽

Immune Correlate ◽

Relative Standard ◽

Limit Of Quantitation

ABSTRACT A Luminex-based direct immunoassay (dLIA) platform has been developed to replace the standardized pneumococcal enzyme-linked immunosorbent assay platform. The multiplex dLIA simultaneously measures the concentration of serum immunoglobulin G (IgG) antibodies specific for pneumococcal capsular polysaccharide (PnPS) serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F. The assay uses poly-l-lysine (PLL)-conjugated PnPS, chemically coupled to spectrally distinct Luminex microspheres. Assay validation experiments were performed using residual human serum samples obtained from 13-valent pneumococcal conjugate vaccine (13vPnC) clinical studies. Assay results are expressed as IgG antibody concentrations in micrograms per milliliter using the international reference serum, 007sp. The lower limit of quantitation (LLOQ) for all serotypes covered in the 13-plex dLIA fell within the range of 0.002 to 0.038 µg/ml serum IgG. The difference between the lower limit and upper limit of the assay range was >500-fold for all serotypes, and assay variability was <20% relative standard deviation (RSD) for all serotypes. IgG antibody measurements were shown to be serotype-specific (some cross-reactivity was observed only between the structurally related serotypes 6A and 6B as well as 19A and 19F), and no interference was observed between the serotypes when the assay was performed in the 13-plex format compared to the singleplex assays. The 13-plex dLIA platform developed by Pfizer Inc. generates up to 143 test results in a single 96-well plate and is a suitable replacement of the enzyme-linked immunosorbent assay (ELISA) platform for evaluating vaccine clinical trials. IMPORTANCE The pneumococcal enzyme-linked immunosorbent assay (ELISA) measures IgG antibodies in human serum, and it is an important assay that supports licensure of pneumococcal vaccines. The immune correlate of protection, 0.35 µg/ml of IgG antibodies, was determined by the ELISA method. Pfizer has developed a new Luminex-based assay platform to replace the ELISA. These papers describe the important work of (i) validating the Luminex-based assay and (ii) bridging the immune correlate of protection (0.35 µg/ml IgG) to equivalent values reported by the Luminex platform.

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Kinetics of IgG Antibodies in Previous Cases of Dengue Fever—A Longitudinal Serological Survey

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph17186580 ◽

2020 ◽

Vol 17 (18) ◽

pp. 6580

Author(s):

Qilin Wu ◽

Qinlong Jing ◽

Xiujuan Wang ◽

Lili Yang ◽

Yilan Li ◽

...

Keyword(s):

Dengue Fever ◽

Antibody Level ◽

Southern China ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibodies ◽

Igg Antibody ◽

Related Factors ◽

Factors Affecting ◽

Serological Investigation ◽

Significant Difference

Guangzhou is believed to be the most important epicenter of dengue outbreaks in southern China. In this study, a longitudinal serological investigation of previous cases of dengue fever in Guangzhou was conducted to explore the persistence of IgG antibodies and related factors affecting the changes of antibody level. We recruited 70 dengue virus type 1 (DENV-1) primary infection cases at two years post infection for serological investigation and conducted a second follow-up in the 5th year of prognosis. An enzyme-linked immunosorbent assay (ELISA) for DENV IgG antibody was examined in all study subjects. Potential factors associated with the concentration of serum total IgG antibody were determined by the generalized estimation equation (GEE). No significant difference in serum total IgG antibody positive rate between two follow-ups was observed (χ2 = 3.066, p = 0.080). However, there was a significant difference in the concentration of serum total IgG antibody between the two follow-ups (Z = 7.154, p < 0.001). The GEE showed that the antibody level in the five-year prognosis was mainly affected by the antibody level in the two-year prognosis (OR: 1.007, 95%CI: 1.005–1.009). In conclusion, the serum IgG antibodies of previous dengue fever cases can persist for a long time.

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Otitis Media in the Young Infant: An IgE-Mediated Disease?

Annals of Otology Rhinology & Laryngology ◽

10.1177/00034894800890s334 ◽

1980 ◽

Vol 89 (3_suppl) ◽

pp. 133-137 ◽

Cited By ~ 8

Author(s):

John L. Sloyer ◽

John H. Ploussard ◽

Laurel J. Karr

Keyword(s):

Otitis Media ◽

Otitis Media With Effusion ◽

Skin Testing ◽

Young Infant ◽

Specific Ige ◽

Serum Samples ◽

Igg Antibody ◽

Total Ige ◽

Ige Antibody ◽

The Mean

IgE antibody directed against noncapsular antigens of mechanically disrupted Streptococcus pneumoniae, serotype 3 rough, was demonstrated in middle ear effusions (MEE) and serum of infants with and without prior evidence of pneumococcal otitis media with effusion (OME). The techniques employed included radioimmunoassay (RIA), passive skin testing, Prausnitz-Küstner (P/K), and enzyme-linked immunospecific assay (ELISA). Adsorption of MEE with ultrasonically disrupted crude pneumococcal antigen (CPA-U) resulted in a reduction of total IgE counts per minute and suggested bacteria-specific IgE antibody ranging from approximately 22 to 92% of the total IgE. The biological activity of the IgE antibody was confirmed by challenging skin passively sensitized with MEE IgE and CPA-U. Areas of induration appeared 20 minutes after challenge and continued to increase in size until 90 minutes. An ELISA procedure was developed as a tool to determine the nature of the antigen(s) and to determine the class(es) of antibody other than IgE. It appeared that CPA-U possesses free amino groups and that it can withstand the rigors of autoclaving. An analysis of 45 cord bloods revealed that high levels of IgG CPA-U antibody occur in this type of sample and that no correlation exists between the IgE and IgG levels. The mean IgG: IgE ratio, optical density at 420 nm (OD 420), for cord bloods was 2.49. In contrast, serum samples from nine infants without pneumococcal otitis media and from 14 infants with pneumococcal otitis media had lower levels of IgG antibody. There was no significant relationship between IgG and IgE OD 420 in infants who never had an episode of pneumococcal otitis media and the mean IgG: IgE was 1.88, whereas the ratio for those infants with pneumococcal otitis media was 1.56. In addition, there was a significant correlation between IgG and IgE levels in this latter group. The results suggest that it may be important to monitor the levels of at least these two classes of antibody to enhance our understanding of the pathogenesis and recovery from otitis media.

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Pneumococcal Type 22F Polysaccharide Absorption Improves the Specificity of a Pneumococcal-Polysaccharide Enzyme-Linked Immunosorbent Assay

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.8.2.266-272.2001 ◽

2001 ◽

Vol 8 (2) ◽

pp. 266-272 ◽

Cited By ~ 210

Author(s):

Nelydia F. Concepcion ◽

Carl E. Frasch

Keyword(s):

Correlation Coefficient ◽

Immunosorbent Assay ◽

Pneumococcal Vaccination ◽

Correlation Coefficients ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibody ◽

Functional Activities ◽

Antibody Titers ◽

The Common ◽

Elisa Inhibition

ABSTRACT The specificity of the immune response to the 23-valent pneumococcal-polysaccharide (PS) vaccine in healthy adults and to a pneumococcal conjugate vaccine in infants was examined by measuring immunoglobulin G (IgG) antibody titers by enzyme-linked immunosorbent assay (ELISA) and the opsonophagocytosis assay. ELISA measures total antipneumococcal IgG titers including the titers of functional and nonfunctional antibodies, while the opsonophagocytosis assay measures only functional-antibody titers. Twenty-four pairs of pre- and post-pneumococcal vaccination sera from adults were evaluated (ELISA) for levels of IgG antibodies against serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F. Twelve of the pairs were also examined (opsonophagocytosis assay) for their functional activities. The correlation coefficients between assay results for most types ranged from 0.75 to 0.90, but the correlation coefficient was only about 0.6 for serotypes 4 and 19F. The specificities of these antibodies were further examined by the use of competitive ELISA inhibition. A number of heterologous polysaccharides (types 11A, 12F, 15B, 22F, and 33A) were used as inhibitors. Most of the sera tested showed cross-reacting antibodies, in addition to those removed by pneumococcal C PS absorption. Our data suggest the presence of a common epitope that is found on most pneumococcal PS but that is not absorbed by purified C PS. Use of a heterologous pneumococcal PS (22F) to adsorb the antibodies to the common epitope increased the correlation between the IgG ELISA results and the opsonophagocytosis assay results. The correlation coefficient improve from 0.66 to 0.92 for type 4 and from 0.63 to 0.80 for type 19F. These common-epitope antibodies were largely absent in infants at 7 months of age, suggesting the carbohydrate nature of the epitope.

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Immune complexes containing factor V in a patient with an acquired neutralizing antibody

Blood ◽

10.1182/blood.v65.4.810.810 ◽

1985 ◽

Vol 65 (4) ◽

pp. 810-818 ◽

Cited By ~ 20

Author(s):

HC Chiu ◽

AK Rao ◽

C Beckett ◽

RW Colman

Keyword(s):

Immunosorbent Assay ◽

Immune Complexes ◽

Neutralizing Antibody ◽

Circulating Immune Complexes ◽

Heat Inactivation ◽

Enzyme Linked Immunosorbent Assay ◽

Antibody Concentration ◽

Factor V ◽

Igg Antibody ◽

Plasma Factor

Abstract An 82-year-old woman presented with extensive hematomas and melena associated with markedly decreased plasma factor V coagulant activity (FV:C). Using a competitive enzyme-linked immunosorbent assay developed in our laboratory, we made serial measurements of factor V antigen (FV:Ag) in plasma and found it to be normal or elevated. The patient's plasma was demonstrated to contain an IgG antibody that could neutralize FV:C in normal plasma. The antibody was of restricted heterogeneity (IgG1, IgG2,kappa). Circulating immune complexes containing antibody to factor V and FV:Ag were demonstrated directly in the plasma by immunoelectrophoresis with polyclonal monospecific antibody and with a monoclonal antibody using an enzyme-linked immunosorbent assay. Presence of neutralizing antibody could be demonstrated in vitro even at times when FV:C was within normal limits by heat inactivation of FV:C. Treatment with plasma and platelet transfusions as well as plasmapheresis induced definite but transient elevation of FV:C. Steroid therapy lowered the neutralizing antibody concentration and produced a rapid and persistent elevation of FV:C during two separate hospitalizations. This report describes a patient in whom levels of FV:Ag have been serially measured, and the presence of circulating immune complexes consisting of factor V and a neutralizing antibody have been directly demonstrated.

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