Synthese und Eigenschaften fluoreszierender Estradiol‐Derivate

Mühlenbruch, Brigitte; Kirmeier, Friedhelm; Roth, H. J.

doi:10.1002/ardp.19863190303

Cited by 16 publications

(5 citation statements)

References 7 publications

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“…This is in agreement with other direct quenching fluoroimmunoassays with small haptens where the fluorescence of the tracer is only partially quenched upon Ab binding. 11, 54 In fact, complete quenching (100%) has only been reported on biotin-avidin systems characterized by even a stronger binding than the Ag-Ab; it has been attributed to the very tight association that brings avidin to the surface of the biotin-fluorophore conjugate. 55 It can be anticipated that a hapten with a shorter spacer arm on the fluorescent label would have given rise to a more efficient quenching due to closer proximity of the fluorophore to the Ab in the Ab-Ag complex.…”

Section: Resultsmentioning

confidence: 99%

“…In particular, fluorescein has an effective excitation wavelength range of about 488−495 nm, closely matching the emission of an Ar laser (488 nm). Frequently, fluoresceinamine is used for labeling haptens containing a COOH group, employing dicyclohexylcarbodiimide (DCC) as an activating agent to form the amide bond. ,, In our case, hapten A was coupled to fluoresceinamine using DIC in THF containing 6% DMF. This activating agent offers the advantage of allowing elimination of the nonreacted DIC and the diisopropylurea byproduct while the solvent evaporates, facilitating subsequent purification steps of the product, AF.…”

Section: Resultsmentioning

confidence: 99%

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Competitive Quenching Fluorescence Immunoassay for Chlorophenols Based on Laser-Induced Fluorescence Detection in Microdroplets

et al. 2002

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An improved biomonitoring system for the analysis of 2,4,6-trichlorophenol (TCP) in urine samples has been developed. The principle of the biosensor device is the detection of laser-induced fluorescence (LIF) in single microdroplets by a homogeneous quenching fluorescence immunoassay (QFIA). The competitive immunoassay occurs in microdroplets (d = 58,4 microm) produced by a piezoelectric generator system with 10-microm-diameter orifice. A continuous Ar ion laser (488 nm) excites the fluorescent tracer; its fluorescence is detected by a spectrometer attached to a 512 x 512 cooled, charge-coupled device camera. Fluorescence is quenched by specific binding of TCP polyclonal antibodies to the fluorescent tracer (hapten A-fluorescein); the quenching effect is diminished by the presence of the analyte. Thus, an increase in the signal is produced in a positive dose-dependent manner when TCP is present in the sample. In 10 mM PBS buffer, the IC50 of the LIF-microdroplet QFIA is 0.45 microg L(-1) reaching a LOD of 0.04 microg L(-1). The QFIA with the same reagents performed in microtiter plate format achieved a LOD of 0.36 microg L(-1) in buffer solution. Performance in human urine was similar to that observed in the buffer. A LOD of 1.6 ,g L(-1), with a dynamic range between 4 and 149.5 microg L(-1) in urine, was obtained without any sample treatment other than dilution with the assay buffer. The detectability achieved is sufficient for occupational exposure risk assessment.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Competitive Quenching Fluorescence Immunoassay for Chlorophenols Based on Laser-Induced Fluorescence Detection in Microdroplets

et al. 2002

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“…From this compound, with one more step, C-7α-biotinylated E 2 affinity ligands can be readily synthesized. It is of note that this intermediate is not just limited to the synthesis of biotinylated E 2 affinity ligands, it can also be used for the synthesis of other useful chemicals such as fluorescent probes for estrogenbinding proteins [19].…”

Section: Chemical Synthesismentioning

confidence: 99%

Chemical synthesis and biochemical characterization of a biotinylated derivative of 17β-estradiol with a long side chain covalently attached to its C-7α position

Jiang

Wang

et al. 2008

Steroids

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High-affinity biotinylated derivatives of 17beta-estradiol (E2) are of value for isolation of various estrogen-binding proteins (including estrogen receptors) and also for studying protein-protein interactions involving these proteins. In this study, we developed a simplified route for the chemical synthesis of a biotinylated derivative of E2 (compound 7) with a side chain attached to its C-7alpha position. Compound 7, i.e., 7alpha-{7-[8-(biotinamido)-octanamido]-heptyl}-estradiol, could be readily synthesized from 6-keto-estradiol-3,17beta-di-tetrahydropyranyl ether (compound 2, which can be prepared from E2), with a final yield of 36%. In vitro receptor-binding assay confirmed that the synthesized affinity ligand has a high binding affinity for both human estrogen receptor alpha and beta. When the affinity ligand (compound 7) was immobilized with avidin on an affinity column, it effectively bound human estrogen receptor alpha, and the receptor protein could be selectively eluted with a biotin-containing buffer. Using the same affinity ligand, prolyl 4-hydroxylase beta-subunit (also known as protein disulfide isomerase) was identified as one of the high-affinity E2-binding proteins in the whole cytosolic protein mixture prepared from MCF-7 human breast cancer cells. Computational molecular modeling analysis showed that compound 7 can bind to human estrogen receptor alpha in a similar manner as ICI-182,780 and raloxifene, and their binding energy values are also similar.

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“…Brassinosteroids (163), comprising a wide range of acid derivatives prepared by esterification or amination of the acid chloride, show growth-promoting activity on a par with that of 24-epi-brassinolide. 141 The pregnadiene (167) gives the dinorcholene (168) in 70 YO yield after treatment with tributyltin hydride in the presence of a,a'-azobisisobutyronitrile.…”

Section: Cholanes Norchoianes and Dinorcholanesmentioning

confidence: 99%

Steroids: reactions and partial syntheses

Turner¹

1988

Nat. Prod. Rep.

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Synthese und Eigenschaften fluoreszierender Estradiol‐Derivate

Cited by 16 publications

References 7 publications

Competitive Quenching Fluorescence Immunoassay for Chlorophenols Based on Laser-Induced Fluorescence Detection in Microdroplets

Competitive Quenching Fluorescence Immunoassay for Chlorophenols Based on Laser-Induced Fluorescence Detection in Microdroplets

Chemical synthesis and biochemical characterization of a biotinylated derivative of 17β-estradiol with a long side chain covalently attached to its C-7α position

Steroids: reactions and partial syntheses

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