A new europium chelate for protein labelling and time-resolved fluorometric applications

Evangelista, Ramon A.; Pollak, Alfred; Allore, Brian; Templeton, Ave F.; Morton, Robert C.; Diamandis, Eleftherios P.

doi:10.1016/0009-9120(88)90006-9

Cited by 179 publications

(86 citation statements)

References 13 publications

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“…Assays based on TRF use lanthanide chelate labels with unique fluorescence properties (24,25,33,34,55,56,58,78,79). Key among these properties is a very long fluorescence decay time and an exceptionally large Stokes' shift.…”

Section: Trf Assaysmentioning

confidence: 99%

Immunological Methods for Detection and Identification of Infectious Disease and Biological Warfare Agents

Peruski

2003

Clin Vaccine Immunol

195

119

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NATURE OF THE PROBLEMBW agents. The release of a biological weapon (BW) agent by a terrorist group or military force would likely be silent and undetectable or nearly so. As shown by anthrax attack during the fall of 2001 in the eastern United States, patients would begin appearing at hospitals and clinics within several days of exposure, most presenting with nonspecific flu-like symptoms. The first days of the outbreak might not even cause undue concern. However, depending on the type of agent and the method of dispersal, the public healthcare system would rapidly be stretched to capacity and beyond.The qualities that make a good BW agent are its relationship between aerosolization, infectivity, or toxicity and the amount of agent required to produce an effect (48). In addition, criteria such as environmental stability, ease of production, disease severity, and communicability determine which agents are the most likely to be utilized. For maximum effect, an optimal agent should be highly lethal and easily produced in large quantities and have limited options for preventive or prophylactic treatment. Given that the respiratory route is the most effective for most BW agents, stability in an aerosol form and the capability to be readily dispersed also in an aerosol (1-to 10-m particle size) are necessary. When potential agents are reviewed for these characteristics, Bacillus anthracis (anthrax) and variola major virus (smallpox) are considered to have the greatest potential for mass casualties and civil disruption. Also high on a prospective list of agents are botulinum neurotoxins, Yersinia pestis, and Francisella tularensis (48,91,92). Lower on the prospective list are Burkholderia pseudomallei and Burkholderia mallei, Rickettsia sp., Coxiella burnetii, Venezuelan equine encephalitis virus, Marburg and Ebola viruses, and influenza viruses (48,63,91,92).Emerging infectious disease agents. In addition to diseases caused by intentional epidemics, there are several emerging infectious diseases (ID) with the potential for significant public health consequences, including dengue fever, West Nile fever, and Rift Valley fever as well as the recent reemergence of malaria in the eastern United States (48,63,91,92). As with BW agents, emerging ID agents may be directly transmissible or vector borne (63). A complex interplay of factors can influence disease emergence, including genetic variation, environmental changes, and population pressures. Further compounding this already complicated situation, are the estimated 600 million international tourists annually, many with the potential to the spread disease globally in a matter of hours (63). Clearly, the challenges facing modern clinical microbiologists and immunologists are daunting enough without the added difficulties posed by the intentional release of BW/ID agents! Because of the threat posed by both BW and emerging or reemerging ID agents, there is a need to rapidly identify such agents in the clinical setting in order to treat the individuals at risk and to improve p...

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Section: Trf Assaysmentioning

confidence: 99%

Immunological Methods for Detection and Identification of Infectious Disease and Biological Warfare Agents

Peruski

2003

Clin Vaccine Immunol

195

119

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“…On the other hand, a hapten-BSA conjugate is widely used to prepare antibodies of the hapten. A BSA molecule has 59 amino groups 21 , some of which are used for binding to the hapten but others still remain unbound, and can be used for binding to BHHCT. In the present experiment, a conjugate (BSM) n (BSA)(BHHCT) 11 (n is unknown) was prepared with a simple and convenient method.…”

Section: Labeling Of the Bsm-bsa Conjugate With Bhhctmentioning

confidence: 99%

Highly Sensitive Detection of Bensulfuron-methyl by Time-Resolved Fluoroimmunoassay Using a Tetradentate β-Diketonate Europium Chelate as a Label

et al. 1999

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A tetradentate β-diketonate europium fluorescent chelate, 4,4′-bis(1″,1″,1″,2″,2″,3″,3″-heptafluoro-4″,6″-hexanedion-6″-yl)-chlorosulfo-o-terphenyl(BHHCT)-Eu 3+ , was used as a label for highly sensitive time-resolved fluoroimmunoassay of bensulfuron-methyl (BSM). A BSM-BSA (BSA=bovine serum albumin) conjugate was labeled with BHHCT-Eu 3+ and the labeled BSM-BSA conjugate was used for competitive immunoassay. The assay was carried out in a 96-well microtiter plate coated with the anti-BSM monoclonal antibody. The method gives the detection limit of 24 pg/ml, which is one to two orders of magnitude lower than those of the conventional HPLC and other methods. The RSD of the present method is less than 10%, and the recovery is in the range of 90 -110% for BSM water samples.

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“…A BSA molecule has 59 amino groups, 24 some of which are used for binding to the E3 molecule, while others are used for binding to BHHCT. In the method, the SA-biotin system, which was employed in the E2 analysis, was not used, since the use of the biotinylated E3-BSA conjugate and the SA-biotin system for the competitive reaction did not give any significant signal.…”

Section: Evaluation Of the Immunoassay Methodsmentioning

confidence: 99%

Quantitative Measurement of 17β-Estradiol and Estriol in River Water by Time-Resolved Fluoroimmunoassay

et al. 2002

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A sensitive method for detecting 17β-estradiol (E2) and estriol (E3) in river water has been developed, based on the timeresolved fluoroimmunoassay by using a fluorescent europium chelate label, 4,4′-bis(1″,1″,1″,2″,2″,3″,3″-heptafluoro-4″,6″-hexanedion-6″-yl)-chlorosulfo-o-terphenyl (BHHCT)-Eu 3+ . In the E2 assay, microtiter plates were coated with the E2-bovine serum albumin (BSA) conjugate. The anti-17β-estradiol antibody, the biotinylated goat anti-rabbit IgG antibody and the BHHCT-Eu 3+ labeled streptavidin (SA)-BSA conjugate were used. In the E3 assay, the goat anti-rabbit IgG antibody was coated on a microtiter plate. The anti-estriol antibody and the BHHCT-Eu 3+ labeled E3-BSA conjugate were used. The detection limits for E2 and E3 were 2.3 pg/ml and 4.3 pg/ml, respectively, and the analytical recoveries were 95 -120%. Quantitative measurement of estrogens in river water was carried out for Kanda River (Tokyo, Japan) by using the method. The E2 and E3 levels were 32 pg/ml and 5.5 pg/ml, respectively. The detection limits of the present method are in the same orders of magnitude as those of ELISA for E2, and are 1 -2 orders of magnitude better for E3.

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A new europium chelate for protein labelling and time-resolved fluorometric applications

Cited by 179 publications

References 13 publications

Immunological Methods for Detection and Identification of Infectious Disease and Biological Warfare Agents

Immunological Methods for Detection and Identification of Infectious Disease and Biological Warfare Agents

Highly Sensitive Detection of Bensulfuron-methyl by Time-Resolved Fluoroimmunoassay Using a Tetradentate β-Diketonate Europium Chelate as a Label

Quantitative Measurement of 17β-Estradiol and Estriol in River Water by Time-Resolved Fluoroimmunoassay

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