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a technology of organometallic probes and probes, which is applied in the field of small organometallic probes, can solve the problems of low targeting of gold, affecting the stability of the probe,
Inactive Publication Date: 2005-06-16
HAINFELD JAMES F +2
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This leads to free antibody which competes for antigen sites and lowers targeting of gold.
Furthermore, the shelf life of the conjugates is compromised by this problem.
Accessibility of the probes to internal cell structures, e.g., nuclear proteins, or to cells deeper in a tissue sample, is impeded by these properties.
The sensitivity of these detection schemes is again degraded by problems of aggregation, detachment of antibodies from the gold, and problems with shelf life.
Although the preparation and properties vary for these metal cluster compounds having organic shells, many of these can only be synthesized in low yields, derivatization for use in coupling to biomolecules is expensive in time and effort, and again in low yields, and many of the cluster compounds are degraded rapidly by heat or various chemical reagents.
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Preparation of Fluorescent and Gold Immunoprobes
1. Preparation of Fluorescein-Conjugated Nanoqold Using Fluorescein-Pbosphine
[0034] A tris (aryl) phosphine ligand bearing a single fluorescein substitutent, and a second tris (aryl) phosphine ligand bearing a single primary amine, were mixed with tris (p-N-methylcarboxamidophenyl) phosphine in the ratio 2:1:5. Ninety mg of this ligand mixture in 25 mL of methanol (an estimated twelve-fold molar excess) was added to a solution of freshly prepared Nanogold (product from 0.4 g of gold (I) triphenylphosphine chloride) in dichloromethane (25 mL) and stirred at room temperature overnight.
[0035] The reaction mixture was extracted with 0.02M ammonium acetate with acetic acid, pH 5.8, in 20% isopropanol / water (3×150 mL), then evaporated to dryness, redissolved in DMSO (2 mL) and 0.6M triethylammonium bicarbonate in 20% isopropanol / water. The fluorescein-substituted Nanogold was isolated by gel filtration, using a coarse gel in a large col...
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Abstract
Small organometallic probes comprise a core of metal atoms bonded to organic moieties. The metal atoms are gold, silver, platinum, palladium, or combinations thereof. In one embodiment, a multifunctional organometallic probe comprises a core of metal atoms surrounded by a shell of organic moieties covalently attached to the metal atoms, a fluorescent molecule, e.g., fluorescein, covalently attached to one of the organic moieties, and a targeting molecule, e.g., an antibody, covalently attached to another of the organic moieties.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation of copending U.S. patent application Ser. No. 10 / 093,770, filed Mar. 8, 2002; which is a continuation-in-part of application Ser. No. 09 / 619,343, filed Jul. 19, 2000 and issued as U.S. Pat. No. 6,369,206; which is a divisional of application Ser. No. 09 / 039,601, filed Mar. 16, 1998 and issued as U.S. Pat. No. 6,121,425; which is a continuation-in-part of application Ser. No. 08 / 652,007, filed May 23, 1996 and issued as U.S. Pat. No. 5,728,590; which is a continuation-in-part of application Ser. No. 08 / 282,929, filed Jul. 29, 1994 and issued as U.S. Pat. No. 5,521,289.BACKGROUND OF THE INVENTION [0002] The present invention is directed to small organometallic probes, processes for making the small organometallic probes, and applications of the small organometallic probes. In particular, the small organometallic probes of the present invention are generally less than two microns (2 μm) across, and compri...
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Patent Type & Authority Applications(United States)
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