EP0406473A1
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EP
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Ion capture reagents and methods for performing binding assays
Assignee: Abbott Laboratories
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Filing: 1989-07-31
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Granted: —
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Expires: 2009-07-31
Abstract
Abstract This invention presents novel reagents, separation techniques and assay procedures which allow both the indicator and the capture reagents to be in solution to avoid problems of slowed immunoreaction kinetics. The separation procedure involves a soluble capture reagent, comprising a specific binding member attached to a charged substance, and an insoluble solid phase that is oppositely charged with respect to the capture reagent. A test sample suspected of containing the analyte of interest is mixed with the capture reagent to form a charged capture reagent/analyte complex. The reaction mixture is contacted to the oppositely charged solid phase to attract, attach, and separate the capture reagent/analyte complex from the reaction mixture. With an appropriate indicator reagent, i.e., a second specific binding substance which is conjugated to a label capable of producing a detectable signal, both sandwich and competitive assays can be performed.
Inventors
- 👤 Robert G. Hiltibran
- 👤 Stephen Denham Stroupe
- 👤 Yi-Her Jou
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-24 10:02
CPC Classifications
C; C08; C08G; C08G69/00; C08G69/02; C08G69/08; C08G69/10; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; G01N33/537; G01N33/538; G01N33/543; G01N33/54366; G01N33/575; G01N33/57565
Claims Analysis
Total Claims: 20
Claim 1
Independent Claim
70 words
An activated polymeric anionic molecule, comprising a compound having the formula: wherein n is about 10 to about 500; z is about 1 to about 6; W is selected from the group consisting of H⁺, Na⁺, K⁺, Li⁺, amine salts, and derivatives thereof; and X is a reactive group or a structure having a reactive group that enables the chemical binding of said activated polymer to a specific binding member.
Claim 2
Dependent Claim
70 words
An activated polymeric anionic molecule, comprising a compound having the formula: wherein n is about 10 to about 500; z is about 1 to about 6; W is selected from the group consisting of H⁺, Na⁺, K⁺, Li⁺, amine salts, and derivatives thereof; and X is a reactive group or a structure having a reactive group that enables the chemical binding of said activated polymer to a specific binding member.
Claim 3
Dependent Claim
46 words
The activated polymeric anionic molecule according to Claim 1, wherein X comprises a spacer of about one to about thirty atoms and said reactive group is selected from the group consisting of an amine-reactive moiety, a thiol-reactive moiety, a thiol moiety and a thiol precursor moiety.
Claim 4
Dependent Claim
37 words
The activated polymeric anionic molecule according to Claim 1, wherein said activated polymeric anionic molecule is selected from the group comprising polyglutamic acid, anionic protein or derivitized proteins, anionic polysaccharides, polyaspartic acid, polyacrylic acid, and polyamino acids.
Claim 5
Dependent Claim
26 words
The activated polymeric anionic molecule according to Claim 3, wherein said derivitized protein is derivatized albumin and wherein said anionic polysaccharide is heparin or alginic acid.
Claim 6
Dependent Claim
92 words
A negatively charged capture reagent, comprising the reaction product of: a specific binding member, and b. an activated polymeric anionic molecule having the formula: wherein n is about 10 to about 500; z is about 1 to about 6; W is selected from the group consisting of H⁺, Na⁺, K⁺, Li⁺, amine salts, and derivatives thereof; and X comprises a spacer of about one to about thirty atoms and a reactive group selected from the group consisting of an amine-reactive moiety, a thiol-reactive moiety, a thiol moiety and a thiol precursor moiety.
Claim 7
Dependent Claim
92 words
A negatively charged capture reagent, comprising the reaction product of: a specific binding member, and b. an activated polymeric anionic molecule having the formula: wherein n is about 10 to about 500; z is about 1 to about 6; W is selected from the group consisting of H⁺, Na⁺, K⁺, Li⁺, amine salts, and derivatives thereof; and X comprises a spacer of about one to about thirty atoms and a reactive group selected from the group consisting of an amine-reactive moiety, a thiol-reactive moiety, a thiol moiety and a thiol precursor moiety.
Claim 8
Dependent Claim
44 words
The negatively charged capture reagent according to Claim 5, wherein said specific binding member has at least one reactive moiety selected from the group consisting of an amino group, a thiol group, a thiol precursor and a thiol-reactive group capable of reacting with X.
Claim 9
Dependent Claim
63 words
A negatively charged capture reagent, comprising the reaction products of: a specific binding member having an amine-reactive group, and b. a polymeric anionic molecule having the formula: wherein n is about 10 to about 500; z is about 1 to about 6; W is selected from the group consisting of H⁺, Na⁺, K⁺, Li⁺, amine salts, and derivatives thereof; and X is H.
Claim 10
Dependent Claim
63 words
A negatively charged capture reagent, comprising the reaction products of: a specific binding member having an amine-reactive group, and b. a polymeric anionic molecule having the formula: wherein n is about 10 to about 500; z is about 1 to about 6; W is selected from the group consisting of H⁺, Na⁺, K⁺, Li⁺, amine salts, and derivatives thereof; and X is H.
Claim 11
Dependent Claim
39 words
The negatively charged capture reagent according to any of Claims 5-7, wherein said polymeric anionic molecule is selected from the group consisting of polyglutamic acid, anionic protein or derivitized proteins, anionic polysaccharides, polyaspartic acid, polyacrylic acid, and polyamino acids.
Claim 12
Dependent Claim
26 words
The negatively charged capture reagent according to Claim 8, wherein said derivitized protein is derivatized albumin and wherein said anionic polysaccharide is heparin or alginic acid.
Claim 13
Independent Claim
191 words
A method for detecting an analyte in a test sample, comprising the steps of: a) contacting the analyte with a capture reagent, comprising a hapten conjugated to a first charged substance, and an indicator reagent, comprising a specific binding member conjugated to a label capable of producing a detectable signal, thereby forming a reaction mixture, wherein said capture reagent is capable of binding a member selected from the group consisting of the analyte, an ancillary specific binding member, said indicator reagent and a complex thereof, and wherein said indicator reagent is capable of binding a member selected from the group consisting of the analyte, an ancillary specific binding member, said capture reagent and a complex thereof, b) contacting said reaction mixture with a solid phase having an opposite charge with respect to said first charged substance, whereby said solid phase attracts and attaches to said first charged substance, thereby enabling the separation of said capture reagent and complexes thereof from said reaction mixture; and c) detecting said label associated with said solid phase or said reaction mixture as an indication of the presence or amount of the analyte in the sample.
Claim 14
Independent Claim
191 words
A method for detecting an analyte in a test sample, comprising the steps of: a) contacting the analyte with a capture reagent, comprising a hapten conjugated to a first charged substance, and an indicator reagent, comprising a specific binding member conjugated to a label capable of producing a detectable signal, thereby forming a reaction mixture, wherein said capture reagent is capable of binding a member selected from the group consisting of the analyte, an ancillary specific binding member, said indicator reagent and a complex thereof, and wherein said indicator reagent is capable of binding a member selected from the group consisting of the analyte, an ancillary specific binding member, said capture reagent and a complex thereof, b) contacting said reaction mixture with a solid phase having an opposite charge with respect to said first charged substance, whereby said solid phase attracts and attaches to said first charged substance, thereby enabling the separation of said capture reagent and complexes thereof from said reaction mixture; and c) detecting said label associated with said solid phase or said reaction mixture as an indication of the presence or amount of the analyte in the sample.
Claim 15
Dependent Claim
32 words
The method according to Claim 10, wherein said capture reagent comprises a specific binding member conjugated to a first charged substance, and said indicator reagent comprises a hapten conjugated to a label
Claim 16
Dependent Claim
31 words
The method according to Claim 10 or 11, wherein a second charged substance having an opposite charge with respect to said first charged substance is retained on said solid phase material.
Claim 17
Dependent Claim
26 words
The method according to Claim 12, wherein said first charged substance is a polymeric anionic substance and said second charged substance is a polymeric cationic substance.
Claim 18
Dependent Claim
27 words
The method according to Claim 13, wherein said polymeric cation is selected from the group consisting of a GAFQuat, diethylaminoethyl-dextran, diallyldimethylammonium chloride-hydroxyethyl cellulose polymer, and hexadimethrine bromide.
Claim 19
Dependent Claim
34 words
The method according to Claim 13, wherein said polymeric anionic molecule is selected from the group consisting of polyglutamic acid, anionic protein or derivitized proteins, anionic polysaccharides, polyaspartic acid, polyacrylic acid, and polyamino acids.
Claim 20
Dependent Claim
23 words
The method according to Claim 15, wherein said derivitized protein is derivatized albumin and wherein said anionic polysaccharide is heparin or alginic acid.
References Cited (Backward Citations)
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