Time-resolved fluorescence of a new europium-chelate complex: demonstration of highly sensitive detection of protein and DNA samples

1993 ◽  
Vol 115 (23) ◽  
pp. 11032-11033 ◽  
Author(s):  
Ashis K. Saha ◽  
Kathryn Kross ◽  
Edmond D. Kloszewski ◽  
Donald A. Upson ◽  
John L. Toner ◽  
...  
Keyword(s):  
Chelate Complex ◽  
Sensitive Detection ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Highly Sensitive ◽  
Europium Chelate ◽  
Highly Sensitive Detection

Eu3+‐labeled IgG‐based time‐resolved fluoroimmunoassay for highly sensitive detection of aflatoxin B1in feed

2017 ◽  
Vol 98 (2) ◽  
pp. 674-680 ◽  
Author(s):  
Xiaofei Hu ◽  
Jingjing Yao ◽  
Fangyu Wang ◽  
Mengqi Yin ◽  
Yaning Sun ◽  
...  
Keyword(s):  
Sensitive Detection ◽  
Time Resolved ◽  
Highly Sensitive ◽  
Highly Sensitive Detection

Ultrasensitive Thyrotropin Immunoassay Based on Enzymatically Amplified Time-Resolved Fluorescence with a Terbium Chelate

1992 ◽  
Vol 38 (4) ◽  
pp. 545-548 ◽  
Author(s):  
A Papanastasiou-Diamandi ◽  
T K Christopoulos ◽  
E P Diamandis
Keyword(s):  
Alkaline Phosphatase ◽  
Detection Limit ◽  
Solid Phase ◽  
Thyroid Stimulating Hormone ◽  
Fluorescence Immunoassay ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Highly Sensitive ◽  
Assay Time ◽  
Highly Sensitive Detection

Abstract We describe an ultrasensitive, enzymatically amplified time-resolved fluorescence immunoassay of thyrotropin (thyroid-stimulating hormone) in serum with use of a terbium chelate as the detectable moiety. In this assay, thyrotropin is first simultaneously reacted with a solid-phase (microtiter well) monoclonal antibody and a soluble biotinylated monoclonal detection antibody. After washing, a streptavidin-alkaline phosphatase conjugate is added, followed by another washing. Alkaline phosphatase acts on the substrate 5-fluorosalicyl phosphate (FSAP) to produce 5-fluorosalicylic acid (FSA). FSA, but not FSAP, can then form with Tb3+ and EDTA a highly fluorescent ternary complex of long fluorescence lifetime. This complex is quantified with time-resolved fluorometry. The thyrotropin assay is highly sensitive (detection limit approximately 0.003 milli-int. unit/L when a total assay time of 85 min is used), precise, and accurate. The thyrotropin assay can also be completed in less than 30 min (detection limit 0.013 milli-int. unit/L), thus making this procedure a candidate technology for high-throughput automated analyzers.

Multiple labeling and time-resolvable fluorophores

1991 ◽  
Vol 37 (9) ◽  
pp. 1486-1491 ◽  
Author(s):  
E P Diamandis
Keyword(s):  
Dicarboxylic Acid ◽  
Detection Limits ◽  
Alpha Fetoprotein ◽  
Macromolecular Complex ◽  
Fluorescence Immunoassay ◽  
Time Resolved Fluorescence ◽  
Time Resolved ◽  
Highly Sensitive ◽  
Europium Chelate ◽  
New Time

Abstract A new time-resolved fluorescence immunoassay system involving use of the europium chelate of 4,7-bis(chlorosulfophenyl)-1,10-phenanthroline-2,9-dicarboxylic acid as label is reviewed. This stable chelate by itself is not very fluorescent but, used in multiple labeling strategies, improves the achievable detection limits. By using multiple labeling, streptavidin tailing, and Eu3+ activation, one can create a very stable, easy-to-use reagent that is suitable for devising highly sensitive immunoassays and other biotechnological assays. This reagent, a streptavidin-based macromolecular complex, is able to detect approximately 300,000 molecules (approximately 0.5 amol) of alpha-fetoprotein in a model noncompetitive immunoassay.

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