A highly-sensitive sandwich enzyme-linked immunosorbent assay for detecting soluble interleukin 6 receptor (sIL-6R) in human serum

2015 ◽  
Author(s):  
Pim van der Harst ◽  
Hilde E. Groot ◽  
Pim van der Harst
Keyword(s):  
Human Serum ◽  
Immunosorbent Assay ◽  
Interleukin 6 ◽  
Enzyme Linked Immunosorbent Assay ◽  
Highly Sensitive ◽  
Interleukin 6 Receptor

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

mSphere ◽  
2018 ◽  
Vol 3 (4) ◽  
pp. e00128-18 ◽  
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.

Enzyme-linked immunosorbent assay for cholesteryl ester transfer protein in human serum

1995 ◽  
Vol 240 (1) ◽  
pp. 1-9 ◽  
Author(s):  
Toshitaka Sato ◽  
Masayoshi Fukasawa ◽  
Makoto Kinoshita ◽  
Hiroyuki Arai ◽  
Takao Saeki ◽  
...  
Keyword(s):  
Cholesteryl Ester ◽  
Human Serum ◽  
Immunosorbent Assay ◽  
Cholesteryl Ester Transfer Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Transfer Protein

Identification of the hemoglobin scavenger receptor/CD163 as a natural soluble protein in plasma

Blood ◽  
2002 ◽  
Vol 99 (1) ◽  
pp. 378-380 ◽  
Author(s):  
Holger Jon Møller ◽  
Niels Anker Peterslund ◽  
Jonas Heilskov Graversen ◽  
Søren Kragh Moestrup
Keyword(s):  
Electrophoretic Mobility ◽  
Immunosorbent Assay ◽  
Broad Spectrum ◽  
Interleukin 6 ◽  
Soluble Protein ◽  
Healthy Subjects ◽  
Scavenger Receptor ◽  
Enzyme Linked Immunosorbent Assay ◽  
Myelomonocytic Leukemia ◽  
Healthy Persons

The hemoglobin scavenger receptor (HbSR/CD163) is an interleukin-6– and glucocorticoid-regulated macrophage/monocyte receptor for uptake of haptoglobin-hemoglobin complexes. Moreover, there are strong indications that HbSR serves an anti-inflammatory function. Immunoprecipitation and immunoblotting enabled identification of a soluble plasma form of HbSR (sHbSR) having an electrophoretic mobility equal to that of recombinant HbSR consisting of the extracellular domain (scavenger receptor cysteine-rich 1-9). A sandwich enzyme-linked immunosorbent assay was established and used to measure the sHbSR level in 130 healthy subjects (median, 1.87 mg/L; range, 0.73-4.69 mg/L). To evaluate the sHbSR levels in conditions with increased leukocyte stimulation and proliferation, 140 patients admitted to a hematological department were screened. Several patients, with a broad spectrum of diagnoses, had a level of sHbSR above the range of healthy persons. Patients with myelomonocytic leukemias and pneumonia/sepsis exhibited the highest levels (up to 67.3 mg/L). In conclusion, sHbSR is an abundant plasma protein potentially valuable in monitoring patients with infections and myelomonocytic leukemia.

An enzyme-linked immunosorbent assay for fetal steroid binding protein of human serum

Steroids ◽  
1985 ◽  
Vol 45 (1) ◽  
pp. 31-38 ◽  
Author(s):  
M. Jawed Iqbal ◽  
Alastair Forbes ◽  
Timothy P. Corbishley ◽  
Mark L. Wilkinson ◽  
Roger Williams
Keyword(s):  
Human Serum ◽  
Immunosorbent Assay ◽  
Binding Protein ◽  
Enzyme Linked Immunosorbent Assay ◽  
Steroid Binding ◽  
Steroid Binding Protein

Development of a Highly Sensitive Enzyme-Linked Immunosorbent Assay Based on Polyclonal Antibodies for the Detection of Polychlorinated Dibenzo-p-dioxins

1998 ◽  
Vol 70 (6) ◽  
pp. 1092-1099 ◽  
Author(s):  
Yukio Sugawara ◽  
Shirley J. Gee ◽  
James R. Sanborn ◽  
S. Douglass Gilman ◽  
Bruce D. Hammock
Keyword(s):  
Immunosorbent Assay ◽  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Highly Sensitive
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