US4326008A
Expired / Lapsed
US
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Protein specific fluorescent microspheres for labelling a protein
Assignee: California Institute of Technology
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Filing: 1980-04-25
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Granted: 1982-04-20
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Expires: 2000-04-25
Abstract
Abstract Highly fluorescent, stable and biocompatible microspheres are obtained by copolymerizing an acrylic monomer containing a covalent bonding group such as hydroxyl, amine or carboxyl, for example, hydroxyethylmethacrylate, with an addition polymerizable fluorescent comonomer such as dansyl allyl amine. A lectin or antibody is bound to the covalent site to provide cell specificity. When the microspheres are added to a cell suspension the marked microspheres will specifically label a cell membrane by binding to a specific receptor site thereon. The labeled membrane can then be detected by fluorescence of the fluorescent monomer.
Inventors
- 👤 Alan Rembaum
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-29 19:54
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/58; G01N33/585; Y; Y10; Y10S; Y10S428/00; Y10S428/913; Y10T; Y10T428/00; Y10T428/29; Y10T428/2982; Y10T428/2991; Y10T428/2998; Y10T428/31504; Y10T428/31855
Claims Analysis
Total Claims: 16
Claim 1
Independent Claim
107 words
A protein specific fluorescent reagent composition for labelling a protein comprising discrete microspheres having a diameter below 100 microns containing a covalent bonding group selected from hydroxyl, amino or carboxyl covalently bound to a conjugate of said protein, said polymer comprising the addition interpolymerized reaction product of a monomer mixture consisting essentially of: 20 to 60% by weight of a first mono-unsaturated acrylic monomer containing at least one of said covalent bonding groups; 0.1% to 15% by weight of a fluorescent comonomer addition polymerizable with said first monomer; and 1 to 20% by weight of a polyunsaturated cross-linking agent addition polymerizable with said first monomer or comonomer.
Claim 2
Dependent Claim
107 words
A protein specific fluorescent reagent composition for labelling a protein comprising discrete microspheres having a diameter below 100 microns containing a covalent bonding group selected from hydroxyl, amino or carboxyl covalently bound to a conjugate of said protein, said polymer comprising the addition interpolymerized reaction product of a monomer mixture consisting essentially of: 20 to 60% by weight of a first mono-unsaturated acrylic monomer containing at least one of said covalent bonding groups; 0.1% to 15% by weight of a fluorescent comonomer addition polymerizable with said first monomer; and 1 to 20% by weight of a polyunsaturated cross-linking agent addition polymerizable with said first monomer or comonomer.
Claim 3
Dependent Claim
14 words
A composition according to claim 1 having a diameter of at least 150 angstrom.
Claim 4
Dependent Claim
26 words
A composition according to claim 1 in which the first monomer is selected from acrylamide, hydroxy-lower alkyl acrylates, amino-lower alkyl acrylates, acrylic acid or methacrylic acid.
Claim 5
Dependent Claim
57 words
A composition according to claim 3 in which the first monomer is selected from compounds of the formula: ##STR3## where R 1 is hydrogen or lower alkyl of 1-8 carbon atoms, R 2 is alkylene of 1-12 carbon atoms, Z is OH or ##STR4## where R 3 or R 4 are H, lower alkyl or lower alkoxy.
Claim 6
Dependent Claim
20 words
A composition according to claim 4 in which the first monomer is selected from 2-hydroxyethyl methacrylate, 3-hydroxypropyl-methacrylate, 2-dimethylaminoethylmethacrylate and 2-aminoethylmethacrylate.
Claim 7
Dependent Claim
14 words
A composition according to claim 4 in which the comonomer is Dansyl allyl amine.
Claim 8
Dependent Claim
14 words
A composition according to claim 4 in which the comonomer is Fluorescamine allyl amine.
Claim 9
Dependent Claim
14 words
A composition according to claim 4 in which the comonomer is Fluorescein allyl amine.
Claim 10
Dependent Claim
42 words
A composition according to claim 1 in which the cross-linking agent is a liquid, polyvinyl diene or triene capable of addition polymerization with the first monomer and comonomer and is present in the monomer mixture in an amount from 5-15% by weight.
Claim 11
Dependent Claim
24 words
A composition according to claim 9 in which the crosslinking agent is selected from ethylene glycol dimethacrylate, divinyl benzene, trimethylol propane trimethacrylate, and N,N'methylene-bis-acrylamide.
Claim 12
Dependent Claim
16 words
A composition according to claim 1 in which the comonomer contains an allyl or vinyl group.
Claim 13
Dependent Claim
36 words
A composition according to claim 1 in which the comonomer consists of the condensation linked reaction product of a fluorochrome containing a functional substitution, A and an allyl or vinyl compound containing a functional substitution, B.
Claim 14
Dependent Claim
33 words
A composition according to claim 12 in which the comonomer is a compound of the formula: Fluorochrome-AB-Olefin where AB is the linking condensation residue selected from urethane, urea, ester, amide, thiourea and sulfonamide.
Claim 15
Dependent Claim
26 words
A composition according to claim 13 in which the fluorochrome A is selected from dansyl chloride, fluorescamine, RB 200 sulfonyl chloride, fluorescein carbonyl chloride and aminofluorescein.
Claim 16
Dependent Claim
25 words
A composition according to claim 14 in which B-Olefin is selected from allyl amine, hydroxy alkyl acrylates, amino alkyl acrylates, methacrylic acid or methacryloyl chloride.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1985-04-19 | 1985-10-19 | 1986-04-20 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1989-04-19 | 1989-10-19 | 1990-04-20 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1993-04-19 | 1993-10-19 | 1994-04-20 | $7700.00 | Window Closed / Lapsed |