Patent US5508164A successfully refreshed from Google Patents.
US5508164A
Expired / Lapsed
US
⏱️ 0.0 years remaining
Isolation of biological materials using magnetic particles
Assignee: DeKalb Genetics Corp
•
Filing: 1993-10-29
•
Granted: 1996-04-16
•
Expires: 2013-10-29
Abstract
Abstract A method for the isolation and sorting of biological materials has been developed. Biological material includes chromosomes, segments of chromosomes, cell organelles, or other minute cellular components. The biological material is separated from the cellular milieu, if necessary, and anchored to a support. Example of a support are glass coverslips, glass or polymer beads. The anchoring is by means of a reversible cross-linking system. The supported biological material is then labelled with compositions capable of binding to said material, and with magnetic particles. Examples of the binding material include nucleic acid probes and antibodies. An example of the antibodies would be those directed to histones. Other labels, for example, fluoresceinbiotin-avidin may be used. The material may be released from the support and sorted by a magnetic force. This method is an alternative to flow cytometry and presents numerous advantages in terms of time, resolution, purity, and preservation of the structure of the biological material during isolation and separation.
Inventors
- 👤 Albert P. Kausch
- 👤 Sandya Narayanswami
- 👤 Jerry E. Manning
- 👤 Barbara A. Hamkalo
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 23:10
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54353; C; C12; C12N; C12N15/00; C12N15/09; C12N15/10; C12N15/1003; C12N15/1006; C12N15/1013; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; C12Q1/6834; G01N33/54313; G01N33/54326; G01N35/00; G01N35/10; G01N35/1009; G01N2035/1025; G01N35/0098
Claims Analysis
Total Claims: 36
Claim 1
Independent Claim
65 words
A method for the magnetic labelling of a chromosome or chloroplast cellular component comprising the following steps: (a) binding said cellular component to a binding composition comprised of an antibody or nucleic acid in the case of chromosome isolation, or antibody in the case of chloroplast isolation; and (b) labelling said binding composition with magnetic particles having an average diameter less than about 2 μm.
Claim 2
Independent Claim
65 words
A method for the magnetic labelling of a chromosome or chloroplast cellular component comprising the following steps: (a) binding said cellular component to a binding composition comprised of an antibody or nucleic acid in the case of chromosome isolation, or antibody in the case of chloroplast isolation; and (b) labelling said binding composition with magnetic particles having an average diameter less than about 2 μm.
Claim 3
Independent Claim
109 words
A method for the isolation of a chromosome or chloroplast cellular component comprising the following steps: (a) reversibly anchoring said cellular component to a support with a DTSSP or SPDP linker; (b) binding said cellular component to a binding composition comprised of an antibody in the case of chromosome or chloroplast isolation, or nucleic acid in the case of chromosome isolation; (c) labelling said binding composition with magnetic particles having an average diameter less than about 2 μm; (d) releasing said cellular component from the support to form free, magnetized cellular components; and (e) isolating said cellular component by applying a magnetic force to said free, magnetized cellular components.
Claim 4
Independent Claim
109 words
A method for the isolation of a chromosome or chloroplast cellular component comprising the following steps: (a) reversibly anchoring said cellular component to a support with a DTSSP or SPDP linker; (b) binding said cellular component to a binding composition comprised of an antibody in the case of chromosome or chloroplast isolation, or nucleic acid in the case of chromosome isolation; (c) labelling said binding composition with magnetic particles having an average diameter less than about 2 μm; (d) releasing said cellular component from the support to form free, magnetized cellular components; and (e) isolating said cellular component by applying a magnetic force to said free, magnetized cellular components.
Claim 5
Dependent Claim
12 words
The method of claim 2, wherein the cellular component is a chromosome.
Claim 6
Dependent Claim
12 words
The method of claim 2, wherein the cellular component is a chloroplast.
Claim 7
Dependent Claim
19 words
The method of claim 2, wherein the binding composition which binds to the cellular component comprises nucleic acid probes.
Claim 8
Dependent Claim
20 words
The method of claim 2, wherein the cellular component comprises polypeptides and the binding composition comprises antibodies to the polypeptides.
Claim 9
Dependent Claim
13 words
The method of claim 2, wherein the magnetic particles comprise ferric oxide particles.
Claim 10
Dependent Claim
35 words
The method of claim 2, wherein the support comprises a silane treated surface that anchors the cellular component to the support, and a cleavable cross-linking agent which crosslinks between the cellular component and the support.
Claim 11
Dependent Claim
15 words
The method of claim 8 wherein the surface is treated with the silane N-(2-aminoethyl)-3-aminopropyl trimethoxysilane.
Claim 12
Dependent Claim
14 words
The method of claim 8 wherein the cleavable cross-linking agent comprises SPDP or DTSSP.
Claim 13
Dependent Claim
12 words
The method of claim 2, wherein the support comprises a glass coverslip.
Claim 14
Dependent Claim
14 words
The method of claim 2, wherein the support comprises controlled pore glass beads (CPG).
Claim 15
Dependent Claim
19 words
The method of claim 12, wherein the controlled pore glass beads are attached to long chain alkylamine linker arms.
Claim 16
Dependent Claim
11 words
The method of claim 2, wherein the support comprises polymer beads.
Claim 17
Dependent Claim
22 words
The method of claim 14, wherein the polymer beads are treated with oligopeptides, said oligopeptides each terminating in a cleavable amino group.
Claim 18
Dependent Claim
12 words
The method of claim 2, wherein the binding composition comprises a label.
Claim 19
Dependent Claim
13 words
The method of claim 16, wherein the label comprises a biotinylated fluorescent marker.
Claim 20
Dependent Claim
23 words
The method of claim 5, wherein the probes are capable of hybridizing to unique repetitive sequences of the centromeric regions of specific chromosomes.
Claim 21
Dependent Claim
19 words
The method of claim 1 wherein the particles are from about 50 nm to about 0.5 microns in diameter.
Claim 22
Dependent Claim
12 words
The method of claim 2, wherein the magnetic particles comprise colloidal gold.
Claim 23
Independent Claim
74 words
A method of separating a chromosome or chloroplast cellular component comprising the following steps: (a) binding a supported, cellular component to a binding composition comprised of an antibody in the case of chromosome or chloroplast isolation, or nucleic acid in the case of chromosome isolation; (b) labelling said bound cellular component with a magnetic particle having an average diameter of less than about 2 μm; and (c) magnetically separating the resulting labelled cellular component.
Claim 24
Independent Claim
74 words
A method of separating a chromosome or chloroplast cellular component comprising the following steps: (a) binding a supported, cellular component to a binding composition comprised of an antibody in the case of chromosome or chloroplast isolation, or nucleic acid in the case of chromosome isolation; (b) labelling said bound cellular component with a magnetic particle having an average diameter of less than about 2 μm; and (c) magnetically separating the resulting labelled cellular component.
Claim 25
Dependent Claim
21 words
The method of claim 21 wherein the cellular component comprises a chromosome and the binding composition comprises a nucleic acid probe.
Claim 26
Dependent Claim
23 words
The method of claim 21 wherein the cellular component comprises a protein and the binding composition comprises an antibody directed to that protein.
Claim 27
Dependent Claim
28 words
The method of claim 21 wherein said cellular component is supported by means mounting said cellular component on a silane treated support by a disulfide-containing bifunctional reversible crosslinker.
Claim 28
Dependent Claim
19 words
The method of claim 21 wherein the particle is from about 50 nm to about 0.5 microns in diameter.
Claim 29
Independent Claim
83 words
A method of sorting chromosomes to achieve purified suspensions thereof, said method comprising the following: (a) anchoring the chromosomes to a support by means of a disulfide-containing bifunctional reversible linker; (b) hybridizing the supported chromosomes with a binding nucleic acid or antibody binding composition to form a complex; (c) labelling the complex with magnetic particles having an average diameter less than about 2 μm; (d) releasing the complex from the support; and (e) sorting the released chromosomes by means of a magnetic force.
Claim 30
Independent Claim
83 words
A method of sorting chromosomes to achieve purified suspensions thereof, said method comprising the following: (a) anchoring the chromosomes to a support by means of a disulfide-containing bifunctional reversible linker; (b) hybridizing the supported chromosomes with a binding nucleic acid or antibody binding composition to form a complex; (c) labelling the complex with magnetic particles having an average diameter less than about 2 μm; (d) releasing the complex from the support; and (e) sorting the released chromosomes by means of a magnetic force.
Claim 31
Dependent Claim
14 words
The method of claim 26, wherein the binding composition comprises nucleic acid sequence probes.
Claim 32
Dependent Claim
11 words
The method of claim 26, wherein the binding composition comprises antibodies.
Claim 33
Independent Claim
79 words
A method of sorting chromosomes to achieve purified preparations thereof, said method comprising the following steps: (a) anchoring the chromosomes to a support by means of a reversible DTSSP or SPDP linker; (b) hybridizing the supported chromosomes with a binding composition comprised of an antibody or nucleic acid to form a complex; (c) labelling the complex with a magnetic label; (d) releasing the complex from the support; and (e) sorting the released chromosomes by means of said magnetic label.
Claim 34
Independent Claim
79 words
A method of sorting chromosomes to achieve purified preparations thereof, said method comprising the following steps: (a) anchoring the chromosomes to a support by means of a reversible DTSSP or SPDP linker; (b) hybridizing the supported chromosomes with a binding composition comprised of an antibody or nucleic acid to form a complex; (c) labelling the complex with a magnetic label; (d) releasing the complex from the support; and (e) sorting the released chromosomes by means of said magnetic label.
Claim 35
Dependent Claim
25 words
The method according to claim 29 wherein said label is a magnetic particle having an average diameter from about 50 nm to about 2 microns.
Claim 36
Dependent Claim
14 words
The method according to claim 29 wherein said label further includes a fluorescent label.
References Cited (Backward Citations)
US3970518A
Cited
US4454234A
Cited
WO1986005815A1
Cited
US4710472A
Cited
US4904391A
Cited
EP0240770A1
Cited
WO1989004373A1
Cited
EP0339980A1
Cited
WO1990006045A2
Cited
WO1990007380A2
Cited
FR2832696A1
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1999-04-16 | 1999-10-16 | 2000-04-16 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2003-04-16 | 2003-10-16 | 2004-04-16 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2007-04-16 | 2007-10-16 | 2008-04-16 | $7700.00 | Window Closed / Lapsed |