Patent US4642284A successfully refreshed from Google Patents.
US4642284A
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Method and system for detection of complement pathway activation
Assignee: Scripps Research Institute
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Filing: 1983-06-13
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Granted: 1987-02-10
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Expires: 2003-06-13
Abstract
Abstract A method and system for detecting and preferably measuring the presence of an activated complement complex in a sample is discussed. The presence of such an activated complex is indicative of complement pathway activation and includes a first complement component and a second complement component. The method uses a first binding agent specific to the first complement component and a second binding agent specific to the second complement component which when bound with the complex forms an aggregate. The second specific binding agent includes a label whose presence is used to detect and measure the amount of aggregate and therefore activated complex in a sample. An assay system and aggregate for use in an assay system are also discussed.
Inventors
- 👤 Neil Cooper
- 👤 James T. Mayes
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-07 02:45
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; G01N33/537; G01N33/564; Y; Y10; Y10S; Y10S435/00; Y10S435/965; Y10S435/966; Y10S435/971; Y10S435/975; Y10S436/00; Y10S436/807; Y10S436/808; Y10S436/821
Claims Analysis
Total Claims: 63
Claim 1
Independent Claim
111 words
An assay method for an activated complex of the complement system comprised of a first complement componment of the complex and second complement component of the complex the assay method to be performed on a sample and comprising the steps of: binding a first specific binding agent to any first complement component forming part of the complex present in the sample; binding a second specific binding agent to any second complement component forming part of the complex, the second specific binding agent including a label, the first and second specific binding agents bound to the complex forming an aggregate; and determining the presence of label bound as part of the aggregate.
Claim 2
Dependent Claim
111 words
An assay method for an activated complex of the complement system comprised of a first complement componment of the complex and second complement component of the complex the assay method to be performed on a sample and comprising the steps of: binding a first specific binding agent to any first complement component forming part of the complex present in the sample; binding a second specific binding agent to any second complement component forming part of the complex, the second specific binding agent including a label, the first and second specific binding agents bound to the complex forming an aggregate; and determining the presence of label bound as part of the aggregate.
Claim 3
Dependent Claim
19 words
The assay method of claim 1 wherein either the first complement component or the second complement component is properdin.
Claim 4
Dependent Claim
13 words
The assay method of claim 1 wherein the first complement component is properdin.
Claim 5
Dependent Claim
16 words
The assay method of claim 3 wherein the first specific binding agent includes an anti-properdin antibody.
Claim 6
Dependent Claim
19 words
The assay method of claim 1 wherein either the first complement component or the second complement component is C3b.
Claim 7
Dependent Claim
13 words
The assay method of claim 1 wherein the second complement component is C3b.
Claim 8
Dependent Claim
16 words
The assay method of claim 6 wherein the second specific binding agent includes an anti-C3b antibody.
Claim 9
Dependent Claim
17 words
The assay method of claim 1 wherein the first specific binding agent includes a functional idiotype-containing polypeptide.
Claim 10
Dependent Claim
17 words
The assay method of claim 1 wherein the second specific binding agent includes a functional idiotype-containing polypeptide.
Claim 11
Dependent Claim
39 words
The assay method of claim 1 including the additional step of separating any second specific binding agent not forming part of the aggregate from the aggregate prior to determining the presence of label bound as part of the aggregate.
Claim 12
Dependent Claim
14 words
The assay method of claim 10 wherein the first specific binding agent is immobilized.
Claim 13
Dependent Claim
47 words
The assay method of claim 11 wherein the sample is first contacted with immobilized first specific binding agent and any portion of the sample which does not bind with the first specific binding agent is removed before the second specific binding agent is bound to the complex.
Claim 14
Dependent Claim
22 words
The assay method of claim 12 wherein any label-containing second specific binding agent not part of the aggregate is removed by washing.
Claim 15
Dependent Claim
36 words
The assay method of claim 10 wherein the first specific binding agent is coupled to a separation means for enhancing the separation of the aggregate from second specific binding agent not forming part of the aggregate.
Claim 16
Dependent Claim
13 words
The assay method of claim 14 wherein the separation means is a carrier.
Claim 17
Dependent Claim
14 words
The assay method of claim 14 wherein the separation means is a solid matrix.
Claim 18
Dependent Claim
20 words
The assay method of claim 1 wherein either the first complement component or the second complement component is factor B.
Claim 19
Dependent Claim
19 words
The assay method of claim 1 wherein either the first complement component or the second complement component is C4b.
Claim 20
Dependent Claim
19 words
The assay method of claim 1 wherein either the first complement component or the second complement component is C2a.
Claim 21
Dependent Claim
30 words
The assay method of claim 1 wherein the first complement component and the second complement components are selected independently from the group consisting of C5b, C6, C7, C8, and C9.
Claim 22
Dependent Claim
24 words
The assay method of claim 1 wherein second specific binding agent not part of the aggregate is separated by a molecular weight-size selective process.
Claim 23
Dependent Claim
12 words
The assay method of claim 1 wherein the label is an enzyme.
Claim 24
Dependent Claim
20 words
The assay method of claim 22 wherein the first specific binding agent is immobilized to form an enzyme-linked immunosorbent assay.
Claim 25
Dependent Claim
29 words
The assay method of claim 1 wherein a predetermined amount of sample is used and the amount of label-including second specific binding agent bound to the aggregate is measured.
Claim 26
Dependent Claim
130 words
An assay method for quantifying complement system activation in a body fluid, the method comprising the steps of: (a) contacting a predetermined amount of body fluid with an immobilized first antibody specific to a first complement component such that the first antibody binds with any first complement component present in the body fluid that forms part of an activated complex including a second complement component, the presence of the complex in the body fluid indicating complement system activation; (b) contacting the complex with a second antibody linked to a label and specific to the second complement component such that the second antibody binds with the second complement component forming part of complex to link the label to the complex; and (c) measuring the quanity of label linked to the complex.
Claim 27
Dependent Claim
130 words
An assay method for quantifying complement system activation in a body fluid, the method comprising the steps of: (a) contacting a predetermined amount of body fluid with an immobilized first antibody specific to a first complement component such that the first antibody binds with any first complement component present in the body fluid that forms part of an activated complex including a second complement component, the presence of the complex in the body fluid indicating complement system activation; (b) contacting the complex with a second antibody linked to a label and specific to the second complement component such that the second antibody binds with the second complement component forming part of complex to link the label to the complex; and (c) measuring the quanity of label linked to the complex.
Claim 28
Dependent Claim
20 words
The assay method of claim 25 including the additional step of removing label not linked to the complex before measuring.
Claim 29
Dependent Claim
19 words
The assay method of claim 26 wherein label-linked second antibody not bound to the complex is removed by washing.
Claim 30
Dependent Claim
23 words
The assay method of claim 25 wherein alternative pathway activation is quantified and presence of the complex is indicative of alternative pathway activation.
Claim 31
Dependent Claim
21 words
The assay method of claim 28 wherein the first complement component is properdin and the first antibody is an anti-properdin antibody.
Claim 32
Dependent Claim
12 words
The assay method of claim 26 wherein the body fluid is serum.
Claim 33
Dependent Claim
12 words
The assay method of claim 26 wherein the label is an enzyme.
Claim 34
Dependent Claim
20 words
The assay method of claim 26 wherein the second complement component is C3b and the second antibody is anti-C3b antibody.
Claim 35
Dependent Claim
25 words
The assay method of claim 26 wherein the body fluid is contacted with the first antibody before the complex is contacted with the second antibody.
Claim 36
Dependent Claim
25 words
The assay method of claim 26 wherein the complex is contacted with the second antibody before the body fluid is contacted with the first antibody.
Claim 37
Dependent Claim
93 words
The assay method of claim 25 further including the steps of providing a known quantity of second complement component immobilized separately from the first antibody, contacting the immobilized second complement component with a separate quantity of the second antibody linked to the label such that the second antibody binds with the immobilized second complement component, measuring the quantity of label linked to the immobilized second complement component as a reference standard, and comparing the quantity of label-linked second antibody bound to the complex with the measured quantity of label of the reference standard.
Claim 38
Dependent Claim
121 words
An assay method for quantifying alternative complement pathway activation in a serum sample comprising the steps of: (a) immobilizing a functional anti-properdin idiotype-containing polypeptide on a solid matrix; (b) linking a labelling enzyme to a functional anti-C3b idiotype-containing polypeptide; (c) contacting a predetermined amount of serum sample with the immobilized anti-properdin polypeptide such that any activated complex comprising properdin and C3b present in the serum sample binds with the anti-properdin polypeptide; (d) contacting the labeled anti-C3b polypeptide with the complex such that the labeled anti-C3b polypeptide binds to the complex and links the labeling enzyme to the complex; (e) removing labelling enzyme not linked to the activated complex; and (f) measuring the amount of labelling enzyme remaining linked to the complex.
Claim 39
Dependent Claim
121 words
An assay method for quantifying alternative complement pathway activation in a serum sample comprising the steps of: (a) immobilizing a functional anti-properdin idiotype-containing polypeptide on a solid matrix; (b) linking a labelling enzyme to a functional anti-C3b idiotype-containing polypeptide; (c) contacting a predetermined amount of serum sample with the immobilized anti-properdin polypeptide such that any activated complex comprising properdin and C3b present in the serum sample binds with the anti-properdin polypeptide; (d) contacting the labeled anti-C3b polypeptide with the complex such that the labeled anti-C3b polypeptide binds to the complex and links the labeling enzyme to the complex; (e) removing labelling enzyme not linked to the activated complex; and (f) measuring the amount of labelling enzyme remaining linked to the complex.
Claim 40
Dependent Claim
24 words
The assay method of claim 36 wherein the sample is contacted with the anti-properdin polypeptide before the anti-C3b polypeptide is contacted with the complex.
Claim 41
Dependent Claim
83 words
The assay method of claim 36 further comprising the steps of immobilizing a known quantity of C3b separately from the anti-properdin polypeptide, contacting a separate quantity of labeled anti-C3b polypeptide with the immobilized C3b such that the labeled anti-C3b polypeptide binds to the immobilized C3b, removing any labeled anti-C3b polypeptide not bound to the immobilized C3b, measuring the amount of label linked to the immobilized C3b as a reference standard, and comparing the measurement label linked to the complex with the reference standard.
Claim 42
Dependent Claim
111 words
A diagnostic assay system for determining the presence an activated complex of the complement system including a first complement component and a second complement component in a sample, the system comprising in kit form: (a) a first specific binding agent specific to the first complement component; (b) a second specific binding agent specific to the second complement component and including a label; when contacted with the complex in the sample to be assayed, the first and second specific binding agents individually binding to and forming an aggregate with the complex, such that the aggregate so formed is separable from any second specific binding agent that is not part of the aggregate.
Claim 43
Dependent Claim
111 words
A diagnostic assay system for determining the presence an activated complex of the complement system including a first complement component and a second complement component in a sample, the system comprising in kit form: (a) a first specific binding agent specific to the first complement component; (b) a second specific binding agent specific to the second complement component and including a label; when contacted with the complex in the sample to be assayed, the first and second specific binding agents individually binding to and forming an aggregate with the complex, such that the aggregate so formed is separable from any second specific binding agent that is not part of the aggregate.
Claim 44
Dependent Claim
21 words
The assay system of claim 39 further including a solid matrix on which the first specific binding agent can be immobilized.
Claim 45
Dependent Claim
33 words
The assay system of claim 39 including a separation means coupled to the first specific binding agent for enhancing separation of the aggregate from second specific binding agent not part of the aggregate.
Claim 46
Dependent Claim
13 words
The assay system of claim 41 wherein the separation means is a carrier.
Claim 47
Dependent Claim
16 words
The assay system of claim 39 wherein the specific binding agents each include an idiotype-containing polypeptide.
Claim 48
Dependent Claim
16 words
The assay system of claim 43 wherein the second specific binding agent includes an anti-C3b antibody.
Claim 49
Dependent Claim
16 words
The assay system of claim 43 wherein the first specific binding agent includes an anti-properdin antibody.
Claim 50
Dependent Claim
12 words
The assay system of claim 39 wherein the label is an enzyme.
Claim 51
Dependent Claim
118 words
A diagnostic assay system for determining the activation of the complement system by the presence in a body fluid sample of an activated complex including a first complement component and a second complement component, the system comprising in kit form: (a) a first antibody to the first complement component of the complex coupled to a solid matrix; and (b) a solution containing a second antibody to the second complement component of the complex and linked to a label; when contacted with the complex, the coupled first antibody binding the first complement component of the complex and the second, label-linked antibody binding to the second complement component of the complex thereby forming an aggregate coupled to the solid matrix.
Claim 52
Dependent Claim
118 words
A diagnostic assay system for determining the activation of the complement system by the presence in a body fluid sample of an activated complex including a first complement component and a second complement component, the system comprising in kit form: (a) a first antibody to the first complement component of the complex coupled to a solid matrix; and (b) a solution containing a second antibody to the second complement component of the complex and linked to a label; when contacted with the complex, the coupled first antibody binding the first complement component of the complex and the second, label-linked antibody binding to the second complement component of the complex thereby forming an aggregate coupled to the solid matrix.
Claim 53
Dependent Claim
21 words
The assay system of claim 47 further including a separate standard reference having a known amount of immobilized second complement component.
Claim 54
Dependent Claim
14 words
The assay system of claim 47 wherein the second antibody is an anti-C3b antibody.
Claim 55
Dependent Claim
14 words
The assay system of claim 47 wherein the first antibody is an anti-properdin antibody.
Claim 56
Dependent Claim
12 words
The assay system of claim 47 wherein the label is an enzyme.
Claim 57
Dependent Claim
63 words
An aggregate for use in an assay method comprising: (a) an activated complex indicative of activation of the complement system and comprising a first complement component and a second complement component; (b) a first specific binding agent bound to the first component in the complex; and (c) a second specific binding agent including a label bound to the second component in the complex.
Claim 58
Dependent Claim
63 words
An aggregate for use in an assay method comprising: (a) an activated complex indicative of activation of the complement system and comprising a first complement component and a second complement component; (b) a first specific binding agent bound to the first component in the complex; and (c) a second specific binding agent including a label bound to the second component in the complex.
Claim 59
Dependent Claim
9 words
The aggregate of claim 52 wherein complex includes C3b.
Claim 60
Dependent Claim
10 words
The aggregate of claim 52 wherein the complex includes properdin.
Claim 61
Dependent Claim
11 words
The aggregate of claim 52 wherein the label is an enzyme.
Claim 62
Dependent Claim
54 words
An aggregate for use in an assay method for indicating activation of the alternative complement pathway comprising: (a) an activated complex including C3b and properdin; (b) an anti-properdin antibody bound to the properdin in the complex; and (c) a C3b antibody linked to a labelling enzyme and bound to the C3b in the complex.
Claim 63
Dependent Claim
54 words
An aggregate for use in an assay method for indicating activation of the alternative complement pathway comprising: (a) an activated complex including C3b and properdin; (b) an anti-properdin antibody bound to the properdin in the complex; and (c) a C3b antibody linked to a labelling enzyme and bound to the C3b in the complex.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1990-02-09 | 1990-08-11 | 1991-02-10 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1994-02-09 | 1994-08-11 | 1995-02-10 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1998-02-09 | 1998-08-11 | 1999-02-10 | $7700.00 | Window Closed / Lapsed |