Detection of tetanus antitoxin using Eu(3+)-labeled anti-human immunoglobulin G monoclonal antibodies in a time-resolved fluorescence immunoassay.

Abstract We describe a liquid-phase nonseparation time-resolved fluorescence immunoassay for measuring estrone-3-glucuronide in undiluted urine. The sensitivity, specificity, and accuracy are similar to those for a conventional separation fluoroimmunoassay or radioimmunoassay, but the speed, convenience, precision, reliability, and clinical utility of the new method are more advantageous. The labeled antigen, a fluorescent europium chelate covalently linked to estrone-3-glucuronide, is incubated for 10 min with a limited concentration of polyclonal or monoclonal antibodies to estrone-3-glucuronyl-6-bovine serum albumin and 10 microL of standard or sample (undiluted urine) in microtiter wells. The fluorescence emanating from the antibody-free label, which is proportional to the concentration of estrone-3-glucuronide in the standard or sample, is then measured in a time-resolved fluorometer. The method is useful for monitoring ovarian function in women.

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Immunogenic and antigenic epitopes of immunoglobulins: studies using monoclonal antibodies to the C γ 3 domain of human immunoglobulin G*

Biochemical Society Transactions ◽

10.1042/bst0100225 ◽

1982 ◽

Vol 10 (4) ◽

pp. 225-226 ◽

Cited By ~ 2

Author(s):

MOHAMMAD ISKANDAR NIK JAAFAR ◽

ROYSTON JEFFERIS

Keyword(s):

Monoclonal Antibodies ◽

Immunoglobulin G ◽

Human Immunoglobulin ◽

Human Immunoglobulin G ◽

Antigenic Epitopes

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Fluorescence immunoassay based on carbon dots as labels for the detection of human immunoglobulin G

Analytical Methods ◽

10.1039/c4ay00717d ◽

2014 ◽

Vol 6 (12) ◽

pp. 4430-4436 ◽

Cited By ~ 34

Author(s):

Lei Zhu ◽

Xin Cui ◽

Jing Wu ◽

Zhenni Wang ◽

Peiyao Wang ◽

...

Keyword(s):

Quantum Yield ◽

Quantitative Determination ◽

Immunoglobulin G ◽

Carbon Dots ◽

Human Immunoglobulin ◽

High Quantum Yield ◽

Fluorescence Immunoassay ◽

Human Immunoglobulin G ◽

High Quantum

A novel fluorescence immunoassay protocol, using high quantum yield of carbon dots as fluorescent labels, was developed for rapid quantitative determination of the model analyte human immunoglobulin G.

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The effects of carbamylation on the antigenicity of human immunoglobulin G probed with monoclonal antibodies

Biochemical Society Transactions ◽

10.1042/bst0150472 ◽

1987 ◽

Vol 15 (3) ◽

pp. 472-473 ◽

Cited By ~ 1

Author(s):

ROYSTON JEFFERIS ◽

LISA F. HODGSON

Keyword(s):

Monoclonal Antibodies ◽

Immunoglobulin G ◽

Human Immunoglobulin ◽

Human Immunoglobulin G

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The differences in short- and long-term varicella-zoster virus (VZV) immunoglobulin G levels following varicella vaccination of healthcare workers measured by VZV fluorescent-antibody-to-membrane-antigen assay (FAMA), VZV time-resolved fluorescence immunoassay and a VZV purified glycoprotein enzyme immunoassay

Epidemiology and Infection ◽

10.1017/s0950268816000595 ◽

2016 ◽

Vol 144 (11) ◽

pp. 2345-2353 ◽

Cited By ~ 3

Author(s):

P. A. C. MAPLE ◽

J. HAEDICKE ◽

M. QUINLIVAN ◽

S. P. STEINBERG ◽

A. A. GERSHON ◽

...

Keyword(s):

Immunoglobulin G ◽

Healthcare Workers ◽

Fluorescent Antibody ◽

Varicella Vaccination ◽

Fluorescence Immunoassay ◽

Time Resolved Fluorescence ◽

Time Resolved ◽

Varicella Zoster ◽

Antigen Assay

SUMMARYHealthcare workers (HCWs) reporting no history of varicella frequently receive varicella vaccination (vOka) if they test varicella-zoster virus (VZV) immunoglobulin G (IgG) negative. In this study, the utilities of VZV-IgG time-resolved fluorescence immunoassay (VZV-TRFIA) and a commercial VZV-IgG purified glycoprotein enzyme immunoassay (gpEIA) currently used in England for confirming VZV immunity have been compared to the fluorescent-antibody-to-membrane-antigen assay (FAMA). A total of 110 HCWs received two doses of vOka vaccine spaced 6 weeks apart and sera collected pre-vaccination (n = 100), at 6 weeks post-completion of vaccination (n = 86) and at 12–18 months follow-up (n = 73) were analysed. Pre-vaccination, by FAMA, 61·0% sera were VZV IgG negative, and compared to FAMA the sensitivities of VZV-TRFIA and gpEIA were 74·4% [95% confidence interval (CI) 57·9–87·0] and 46·2% (95% CI 30·1–62·8), respectively. Post-completion of vaccination the seroconversion rate by FAMA was 93·7% compared to rates of 95·8% and 70·8% determined by VZV-TRFIA and gpEIA, respectively. At 12–18 months follow-up seropositivity rates by FAMA, VZV-TRFIA and gpEIA were 78·1%, 74·0% and 47·9%, respectively. Compared to FAMA the sensitivities of VZV-TRFIA and gpEIA for measuring VZV IgG following vaccination were 96·4% (95% CI 91·7–98·8) and 74·6% (95% CI 66·5–81·6), respectively. Using both FAMA and VZV-TRFIA to identify healthy adult VZV susceptibles and measure seroconversion showed that vOka vaccination of HCWs is highly immunogenic.

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Performance of a Time-Resolved Fluorescence Immunoassay for Measuring Varicella-Zoster Virus Immunoglobulin G Levels in Adults and Comparison with Commercial Enzyme Immunoassays and Merck Glycoprotein Enzyme Immunoassay

Clinical and Vaccine Immunology ◽

10.1128/cvi.13.2.214-218.2006 ◽

2006 ◽

Vol 13 (2) ◽

pp. 214-218 ◽

Cited By ~ 35

Author(s):

P. A. C. Maple ◽

J. Gray ◽

J. Breuer ◽

G. Kafatos ◽

S. Parker ◽

...

Keyword(s):

Varicella Zoster Virus ◽

Immunoglobulin G ◽

High Specificity ◽

Fluorescence Immunoassay ◽

Time Resolved Fluorescence ◽

Time Resolved ◽

Commercial Enzyme ◽

Varicella Zoster ◽

Enzyme Immunoassays ◽

Human Sera

ABSTRACT Highly sensitive and specific, quantitative assays are needed to detect varicella-zoster virus (VZV) immunoglobulin G in human sera, particularly for determining immune status and response following vaccination. A time-resolved fluorescence immunoassay (TRFIA) has been developed, and its performance was compared to that of two commercial enzyme immunoassays (EIAs) and Merck glycoprotein EIA (gpEIA). The TRFIA had equivalent sensitivity (97.8%) and high specificity (93.5%) in relation to gpEIA. A commercial (Behring) EIA compared favorably with TRFIA in terms of sensitivity (98.4%) but had lower specificity (80.7%). Another commercial EIA (Diamedix) had high specificity (97.1%) but low sensitivity (76.4%) compared to TRFIA if equivocal test results were treated as negative for VZV antibody. A novel feature of the TRFIA was that the cutoff was generated using population mixture modeling and was expressed in mIU/ml, as the assay was calibrated using the British standard VZV antibody.

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