Patent US4935147A successfully refreshed from Google Patents.
US4935147A
Expired / Lapsed
US
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Particle separation method
Assignee: Siemens Healthcare Diagnostics GmbH Germany
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Filing: 1988-10-26
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Granted: 1990-06-19
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Expires: 2008-10-26
Abstract
Abstract A method is disclosed for separating a substance from a liquid medium. The method comprises combining the liquid medium containing the substance with magnetic particles under conditions for non-specific chemical binding of the magnetic particles. Thereafter, the medium is subjected to a magnetic field gradient to separate the particles from the medium. The preferred non-specific binding is achieved as the result of charge interactions between the particles usually by means of a polyionic reagent. The method of the invention has particular application to the separation of cells and microorganisms from aqueous suspensions and also to the determination of an analyte in a sample suspected of containing the analyte. The analyte is a member of a specific binding pair (sbp). The sample is combined in an assay medium with magnetic particles and a sbp member complementary to the analyte. Magnetic or non-magnetic particles capable of specific binding to the analyte or its complementary sbp member must be included in the assay medium. The combination is made under conditions for non-specifically aggregating the magnetic particles or coaggregating the magnetic and non-magnetic particles when non-magnetic particles are present. The assay medium is subjected to a magnetic field gradient to separate the aggregated particles from the medium. Then, the medium or the particles are examined for the presence or amount of the analyte or an sbp member, the binding of which is affected by the presence of the analyte.
Inventors
- 👤 Edwin F. Ullman
- 👤 Vartan E. Ghazarossian
- 👤 Nurith Kurn
- 👤 Litai Weng
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-04 07:05
CPC Classifications
B; B03; B03C; B03C1/00; B03C1/005; B03C1/01; C; C12; C12Q; C12Q1/00; C12Q1/02; C12Q1/24; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; G01N33/537; G01N33/538; G01N33/543; G01N33/54313; G01N33/54326; G01N33/54333
Claims Analysis
Total Claims: 29
Claim 1
Independent Claim
112 words
In a method for separating suspended charged non-magnetic particles from a liquid medium, said method involving (a) coupling said charged non-magnetic particles to like-charged magnetic particles, and (b) separating the coupled particles from said medium by means of a magnetic field gradient, the improvement which comprises non-specifically coupling said charged non-magnetic particles to said like-charged magnetic particles by inclusion in said medium of chemical means for non-specifically coupling said charged non-magnetic particles to said like-charged magnetic particles, said chemical means being selected from the group consisting of polyalkylene amines and their lower alkyl ammonium salts, aminodextrans, chitosan, protamine, positively charged liposomes, polylysine, heparin, dextran sulfate, negatively charged phospholipid vesicles and polycarboxylic acids.
Claim 2
Dependent Claim
112 words
In a method for separating suspended charged non-magnetic particles from a liquid medium, said method involving (a) coupling said charged non-magnetic particles to like-charged magnetic particles, and (b) separating the coupled particles from said medium by means of a magnetic field gradient, the improvement which comprises non-specifically coupling said charged non-magnetic particles to said like-charged magnetic particles by inclusion in said medium of chemical means for non-specifically coupling said charged non-magnetic particles to said like-charged magnetic particles, said chemical means being selected from the group consisting of polyalkylene amines and their lower alkyl ammonium salts, aminodextrans, chitosan, protamine, positively charged liposomes, polylysine, heparin, dextran sulfate, negatively charged phospholipid vesicles and polycarboxylic acids.
Claim 3
Dependent Claim
11 words
The method of claim 1 wherein said non-specific coupling is reversible.
Claim 4
Dependent Claim
13 words
The method of claim 1 wherein said non-specific coupling results from charge interactions.
Claim 5
Independent Claim
91 words
A method of separating suspended charged non-magnetic particles from a liquid medium, which comprises combining in a liquid medium charged non-magnetic particles and like-charged magnetic particles, and chemical means for non-specifically binding said non-magnetic particles and said magnetic particles, said chemical means being selected from the group consisting of polyalkylene amines and their lower alkyl ammonium salts, aminodextrans, chitosan, protamine, positively charged liposomes, polylysine, heparin, dextran sulfate, negatively charged phospholipid vesicles and polycarboxylic acids, and subjecting said medium to a magnetic field gradient to separate said bound particles from said medium.
Claim 6
Independent Claim
91 words
A method of separating suspended charged non-magnetic particles from a liquid medium, which comprises combining in a liquid medium charged non-magnetic particles and like-charged magnetic particles, and chemical means for non-specifically binding said non-magnetic particles and said magnetic particles, said chemical means being selected from the group consisting of polyalkylene amines and their lower alkyl ammonium salts, aminodextrans, chitosan, protamine, positively charged liposomes, polylysine, heparin, dextran sulfate, negatively charged phospholipid vesicles and polycarboxylic acids, and subjecting said medium to a magnetic field gradient to separate said bound particles from said medium.
Claim 7
Dependent Claim
15 words
The method of claim 4 wherein said non-specific binding is the result of charge interactions.
Claim 8
Dependent Claim
19 words
The method of claim 4 wherein said magnetic particles have a diameter of from about 5 to 100 nm.
Claim 9
Dependent Claim
12 words
The method of claim 4 wherein said non-magnetic particles are biological particles.
Claim 10
Dependent Claim
19 words
The method of claim 7 wherein said biological particles are selected from the group consisting of cells and microorganisms.
Claim 11
Dependent Claim
11 words
The method of claim 4 wherein said non-magnetic particles are erythrocytes.
Claim 12
Dependent Claim
12 words
The method of claim 4 wherein said non-magnetic particles are synthetic particles.
Claim 13
Dependent Claim
20 words
The method of claim 10 wherein said non-magnetic particles are selected from the group consisting of liposomes and latex particles.
Claim 14
Dependent Claim
19 words
The method of claim 4 wherein a member of a specific binding pair is bound to said non-magnetic particles.
Claim 15
Dependent Claim
23 words
The method of claim 4 wherein said bound non-magnetic particles are treated with a reagent to reverse their binding to said magnetic particles.
Claim 16
Dependent Claim
18 words
The method of claim 13 wherein said reagent to reverse said binding is a high ionic strength solution.
Claim 17
Dependent Claim
24 words
The method of claim 13 wherein said polyalkylene amine and ammonium salts thereof are selected from the group consisting of polyethyleneimine, polypropyleneimine and polybrene.
Claim 18
Dependent Claim
19 words
The method of claim 15 wherein said polycarboxylic acids are selected from the group consisting of polyacrylate and polyglutamates.
Claim 19
Dependent Claim
19 words
The method of claim 4 wherein said chemical means is selected from the group consisting of polybrene and polyethyleneimine.
Claim 20
Dependent Claim
23 words
The method of claim 4 wherein said non-magnetic particles are liposomes and after separation said bound non-magnetic particles are combined with a detergent.
Claim 21
Dependent Claim
23 words
The method of claim 4 wherein said magnetic particles are present in a magnetic fluid prior to combination with said non magnetic particles.
Claim 22
Dependent Claim
14 words
The method of claim 4 wherein said magnetic particules are coated with a protein.
Claim 23
Independent Claim
88 words
A method for removing cells from whole blood, which comprises combining in a liquid medium a whole blood sample and negatively charged magnetic particles and chemical means for non-specific binding said magnetic particles to said cells, said chemical means being selected from the group consisting of polyalkylene amines and their lower alkyl ammonium salts, aminodextrans, chitosan, protamine, positively charged liposomes, polylysine, heparin, dextran sulfate, negatively charged phospholipid vesicles and polycarboxylic acids, and subjecting said medium to a magnetic field gradient to separate said bound cells from said medium.
Claim 24
Independent Claim
88 words
A method for removing cells from whole blood, which comprises combining in a liquid medium a whole blood sample and negatively charged magnetic particles and chemical means for non-specific binding said magnetic particles to said cells, said chemical means being selected from the group consisting of polyalkylene amines and their lower alkyl ammonium salts, aminodextrans, chitosan, protamine, positively charged liposomes, polylysine, heparin, dextran sulfate, negatively charged phospholipid vesicles and polycarboxylic acids, and subjecting said medium to a magnetic field gradient to separate said bound cells from said medium.
Claim 25
Dependent Claim
24 words
The method of claim 21 wherein said chemical means is selected from a group consisting of polybrene, polyalkyleneimine, aminodextran, chitosan, and positively charged liposomes.
Claim 26
Dependent Claim
13 words
The method of claim 21 wherein said chemical means is polybrene or polyethyleneimine.
Claim 27
Dependent Claim
16 words
The method of claim 21 wherein said magnetic particles are from about 5-100 nm in diameter.
Claim 28
Dependent Claim
21 words
The method of claim 21 wherein said magnetic particles are present in a ferrofluid prior to combination with said blood sample.
Claim 29
Dependent Claim
14 words
The method of claim 25 wherein said magnetic particles are coated with a protein.
References Cited (Backward Citations)
GB2019378A
Cited
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