EP0265244A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Target and background capture methods and apparatus for affinity assays
Assignee: BP Corp North America Inc
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Filing: 1987-10-21
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Granted: —
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Expires: 2007-10-21
Abstract
Abstract Methods, compositions and instruments for performing affinity assays for target molecules include contacting a sample with a retrievable support capable of forming a substantially homogenous dispersion within a sample medium and reversibly binding to the target molecule. The retrievable support is capable of being separated from the sample medium recapturing the target molecule from other constituents of the sample.
Inventors
- 👤 Mark Leo Collins
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-05 09:02
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/68; C12Q1/6813; B; B03; B03C; B03C1/00; B03C1/005; B03C1/01; B03C1/02; B03C1/025; B03C1/031; B03C1/033; B03C1/0335; B03C1/28; B03C1/288; B03C1/30; C12Q1/6816; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/54313; G01N33/54326; B03C2201/00; B03C2201/18; B03C2201/26; G01N35/00; G01N35/0098
Claims Analysis
Total Claims: 49
Claim 1
Independent Claim
142 words
A method for assaying a sample for a target molecule which is a member of a biological binding pair comprising: a. contacting the sample with reagent and a retrievable support under binding conditions wherein the reagent comprises a first antiligand probe and a second antiligand probe, the first and second probes capable of forming a complex with the target molecule in which the probes are bound to the target, the first probe capable of binding to the retrievable support under binding conditions and the second probe includes a label moiety capable of detection; b. substantially separating the retrievable support from the sample forming a removal product which in the presence of target in the sample, comprises target, and first and second probes; and c. monitoring the removal product for the presence of the label moiety indicating the presence of the target molecule.
Claim 2
Independent Claim
142 words
A method for assaying a sample for a target molecule which is a member of a biological binding pair comprising: a. contacting the sample with reagent and a retrievable support under binding conditions wherein the reagent comprises a first antiligand probe and a second antiligand probe, the first and second probes capable of forming a complex with the target molecule in which the probes are bound to the target, the first probe capable of binding to the retrievable support under binding conditions and the second probe includes a label moiety capable of detection; b. substantially separating the retrievable support from the sample forming a removal product which in the presence of target in the sample, comprises target, and first and second probes; and c. monitoring the removal product for the presence of the label moiety indicating the presence of the target molecule.
Claim 3
Dependent Claim
30 words
The method of Claim 1 wherein the said target includes a polynucleotide and said first and second probes comprises polynucleotides capable of binding to mutually exclusive portions of the target.
Claim 4
Dependent Claim
11 words
The method of Claim 1 wherein said retrievable support comprises beads.
Claim 5
Dependent Claim
16 words
The method of Claim 3 wherein said beads are capable of interacting with a magnetic field.
Claim 6
Dependent Claim
18 words
The method of Claim 1 wherein said retrievable support is capable of substantially homogeneous dispersion within a sample.
Claim 7
Dependent Claim
71 words
The method of Claim 1 wherein said second probe is further capable of binding to a second support when said second probe is unbound to the target, and said method comprises contacting said sample with said second support under binding conditions to allow formation of a complex between said second support and said second probe, and separating said second support from the sample to remove unbound second probe to reduce background.
Claim 8
Independent Claim
44 words
A method of target capture for target polynucleotide molecules comprising contacting a sample potentially containing target polynucleotides with a retrievable bead capable of forming a substantially homogeneous dispersion within said sample medium and said retrievable bead capable of specifically associating with the target polynucleotide.
Claim 9
Independent Claim
44 words
A method of target capture for target polynucleotide molecules comprising contacting a sample potentially containing target polynucleotides with a retrievable bead capable of forming a substantially homogeneous dispersion within said sample medium and said retrievable bead capable of specifically associating with the target polynucleotide.
Claim 10
Dependent Claim
22 words
The method of Claim 7 wherein said bead is capable of associating with said target polynucleotide by means of a probe polynucleotide.
Claim 11
Dependent Claim
21 words
The method of Claim 8 wherein said probe polynucleotide is capable of associating with the retrievable support and the target molecule.
Claim 12
Dependent Claim
12 words
The method of Claim 7 wherein said bead is comprised of polystyrene.
Claim 13
Dependent Claim
17 words
The method of Claim 7 wherein said retrievable bead is capable of interacting with a magnetic field.
Claim 14
Dependent Claim
38 words
The method of Claim 7 further including the step of contacting said sample with a second polynucleotide probe capable of associating with a portion of the target exclusive of said first probe, said second probe capable of detection.
Claim 15
Dependent Claim
43 words
The method of Claim 7 further comprising the step of separating the retrievable support from the sample, contacting the retrievable support with a second medium and subjecting the second medium to releasing conditions to allow release of the target from the retrievable support.
Claim 16
Dependent Claim
39 words
The method of Claim 13 including the additional step of contacting the second medium with a second support, said second support capable of forming a complex with the target under binding conditions to allow capture of the target molecule.
Claim 17
Dependent Claim
37 words
The method of Claim 14 further comprising the step of contacting the second retrievable support with a third medium and subjecting the third medium to releasing conditions to allow release of the target from the retrievable support.
Claim 18
Dependent Claim
32 words
The method of Claim 15 further comprising the step of contacting the third medium with a support capable of forming a complex with the target under binding conditions to recapture the target.
Claim 19
Dependent Claim
26 words
The method of Claim 15 wherein said releasing conditions in said second medium and said releasing conditions in said third medium are physically and chemically distinct.
Claim 20
Dependent Claim
27 words
The method of Claim 17 wherein said releasing conditions in said second medium is selected from the group of releasing methods including chemical elution and thermal elution.
Claim 21
Dependent Claim
14 words
The method of Claim 14 wherein said first and second support are chemically distinct.
Claim 22
Dependent Claim
49 words
A kit for capturing target polynucleotide comprising a retrievable support capable of forming substantially homogenous dispersions within a sample medium, and capable of separation from said sample medium and said kit including a first probe capable of binding to the retrievable support and said target molecule under binding conditions.
Claim 23
Dependent Claim
49 words
A kit for capturing target polynucleotide comprising a retrievable support capable of forming substantially homogenous dispersions within a sample medium, and capable of separation from said sample medium and said kit including a first probe capable of binding to the retrievable support and said target molecule under binding conditions.
Claim 24
Dependent Claim
39 words
The kit of Claim 20 further comprising a second probe capable of binding to the target molecule to form a complex with the target molecule and said first probe, said second probe including a label moiety capable of detection.
Claim 25
Dependent Claim
37 words
A kit for capturing target polynucleotides comprising a retrievable bead capable of forming substantially homogeneous dispersions within a sample medium, said bead capable of specifically binding to said target and capable of separation from said sample medium.
Claim 26
Dependent Claim
37 words
A kit for capturing target polynucleotides comprising a retrievable bead capable of forming substantially homogeneous dispersions within a sample medium, said bead capable of specifically binding to said target and capable of separation from said sample medium.
Claim 27
Dependent Claim
15 words
The kit of Claim 22 wherein said bead is capable of interacting with magnetic fields.
Claim 28
Dependent Claim
57 words
An instrument for performing assays for target polynucleotides comprising a reaction chamber adapted for receiving target polynucleotides in a sample medium and a support capable of substantially homogeneous dispersion within the sample medium and capable of forming a complex with the target, and means for separating the support with the target bound thereto from the sample medium.
Claim 29
Dependent Claim
57 words
An instrument for performing assays for target polynucleotides comprising a reaction chamber adapted for receiving target polynucleotides in a sample medium and a support capable of substantially homogeneous dispersion within the sample medium and capable of forming a complex with the target, and means for separating the support with the target bound thereto from the sample medium.
Claim 30
Dependent Claim
18 words
The instrument of Claim 24 further comprising magnetic means for separating the retrievable support from the sample medium.
Claim 31
Dependent Claim
20 words
The instrument of Claim 25 further including means for releasing the target from said retrievable support into a second medium.
Claim 32
Dependent Claim
19 words
The instrument of Claim 26 further comprising means for detecting label moieties carried upon probes bound to said target.
Claim 33
Dependent Claim
32 words
The instrument of Claim 24 wherein said support forms a complex with the target through at least one probe capable of forming a complex with said target and associating with the support.
Claim 34
Dependent Claim
21 words
The instrument of Claim 27 comprising means for associating probes, carrying label moieties which are unbound to target, with a support.
Claim 35
Independent Claim
77 words
A method of assaying a sample for target molecules including the steps of contacting the sample with reagent including a probe under binding conditions, said probe having a label moiety, and capable of forming a complex with said target under binding conditions and capable of forming a complex with a support when said probe is unbound to target, and contacting said sample and reagent with a support capable of binding unbound probe to allow reduction of background.
Claim 36
Independent Claim
77 words
A method of assaying a sample for target molecules including the steps of contacting the sample with reagent including a probe under binding conditions, said probe having a label moiety, and capable of forming a complex with said target under binding conditions and capable of forming a complex with a support when said probe is unbound to target, and contacting said sample and reagent with a support capable of binding unbound probe to allow reduction of background.
Claim 37
Dependent Claim
30 words
The method of Claim 30 wherein said probe includes a homopolymer ligand capable of binding to said support which includes bases sequences which are capable of binding to said target.
Claim 38
Dependent Claim
29 words
The method of Claim 31 wherein said probe releases from said support at a lower temperature when said probe is bound to target than when said probe is unbound.
Claim 39
Dependent Claim
23 words
The method of Claim 31 wherein said ligand includes a tenmer homopolymer ligand sharing at least three consecutive base sequences with said target.
Claim 40
Dependent Claim
16 words
A bead capable of specific association with a target polynucleotide and dispersible within a liquid medium.
Claim 41
Dependent Claim
16 words
A bead capable of specific association with a target polynucleotide and dispersible within a liquid medium.
Claim 42
Dependent Claim
15 words
The bead of Claim 34 wherein said bead is capable of interacting with magnetic fields.
Claim 43
Dependent Claim
15 words
The bead of Claim 34 further comprising oligonucleotide ligands capable of binding to a probe.
Claim 44
Dependent Claim
12 words
The bead of Claim 34 wherein said bead is comprised of polystyrene.
Claim 45
Dependent Claim
14 words
The bead of Claim 34 wherein said bead has a diameter of 1-10 microns.
Claim 46
Dependent Claim
17 words
The bead of Claim 36 wherein said bead has a surface to which homopolymers are covalently bonded.
Claim 47
Dependent Claim
70 words
A reagent composition for use with a first support having a first antiligand comprising a first probe and a second probe, said first probe capable of forming a complex with a target molecule and includes a first ligand capable of binding to said first antiligand and said second probe capable of forming a complex with said target molecule with said first probe and having a label moiety capable of detection.
Claim 48
Dependent Claim
70 words
A reagent composition for use with a first support having a first antiligand comprising a first probe and a second probe, said first probe capable of forming a complex with a target molecule and includes a first ligand capable of binding to said first antiligand and said second probe capable of forming a complex with said target molecule with said first probe and having a label moiety capable of detection.
Claim 49
Dependent Claim
28 words
The reagent composition of Claim 40 wherein said second second probe includes a second ligand moiety capable of binding with a second antiligand carried on a second support.
References Cited (Backward Citations)
No citation records indexed.
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