Claims Analysis
Total Claims: 23
Claim 1
Independent Claim
217 words
In an immunoassay for the determination of an analyte, where the analyte is a member of a specific binding pair consisting of ligand and antiligand and wherein a label is employed which provides a detectible signal, the value of which is affected by the proximity of said antiligand to said label due to the presence of the bulk of said antiligand or a compound conjugated to said antiligand which interacts with said label, wherein said immunoassay comprises combining in an aqueous medium: (1) a sample suspected of containing said analyte, (2) a reacgent comprising ligand or ligand analog and label, wherein said ligand analog is capable of specific binding to said antiligand, and (3) antiligand, when ligand is the analyte or said antiligand is conjugated with said compound which interacts with said label; and determining the value of said detectible signal as compared to said value determined with a sample having a known amount of analyte; the improvement which comprises employing as said reagent discrete colloidal particles comprised of a major portion of lipophilic compounds and a minor portion of label and ligand or ligand analog, wherein said label is covalently conjugated to at least one of said lipophilic compounds, and the ligand or ligand analog are lipophilic or made so by conjunction to a lipophilic compound.
Claim 2
Dependent Claim
217 words
In an immunoassay for the determination of an analyte, where the analyte is a member of a specific binding pair consisting of ligand and antiligand and wherein a label is employed which provides a detectible signal, the value of which is affected by the proximity of said antiligand to said label due to the presence of the bulk of said antiligand or a compound conjugated to said antiligand which interacts with said label, wherein said immunoassay comprises combining in an aqueous medium: (1) a sample suspected of containing said analyte, (2) a reacgent comprising ligand or ligand analog and label, wherein said ligand analog is capable of specific binding to said antiligand, and (3) antiligand, when ligand is the analyte or said antiligand is conjugated with said compound which interacts with said label; and determining the value of said detectible signal as compared to said value determined with a sample having a known amount of analyte; the improvement which comprises employing as said reagent discrete colloidal particles comprised of a major portion of lipophilic compounds and a minor portion of label and ligand or ligand analog, wherein said label is covalently conjugated to at least one of said lipophilic compounds, and the ligand or ligand analog are lipophilic or made so by conjunction to a lipophilic compound.
Claim 3
Dependent Claim
12 words
A method according to claim 1, wherein said lipophilic compound is amphiphilic.
Claim 4
Dependent Claim
14 words
A method according to claim 2, wherein said amphiphilic compound is a phosphatidyl compound.
Claim 5
Dependent Claim
13 words
A method according to claim 1, wherein said label is a fluorescent compound.
Claim 6
Dependent Claim
16 words
A method according to claim 4, wherein said fluorescent compound is conjugated to a phosphatidyl group.
Claim 7
Dependent Claim
16 words
A method according to claim 5, wherein said ligand is cardiolipin and said analyte is anticardiolipin.
Claim 8
Dependent Claim
227 words
An immunoassay for the determination of an analyte, wherein the analyte is a member of a specific binding pair consisting of ligand and antiligand and wherein a label is employed which provides a detectible signal, the value of which is affected by the proximity of said antiligand to said label due to the presence of the bulk of said antiligand or a compound conjugated to said antiligand which interacts with said label, and wherein a reagent is employed having both label and ligand or a ligand analog, where ligand analog is capable of specifically binding to said antiligand, said reagent comprising a liposome of amphiphilic molecules having from about 0.05 to 15 mole percent of ligand or ligand analog conjugated to an amphiphile and from 0.05 to 15 mole percent of a label conjugated to at least one of said amphiphile molecules; wherein said immunoassay comprises combining in an aqueous medium at a pH in the range of about 5 to 10 and at a temperature in the range of about 10° to 50° C.: a sample suspected of containing said analyte; said reagent; and antiligand, when ligand is the analyte or said antiligand is conjugated with a compound which interacts with said label; and determining the value of said detectible signal as compared to said value determined with a sample having a known amount of analyte.
Claim 9
Dependent Claim
227 words
An immunoassay for the determination of an analyte, wherein the analyte is a member of a specific binding pair consisting of ligand and antiligand and wherein a label is employed which provides a detectible signal, the value of which is affected by the proximity of said antiligand to said label due to the presence of the bulk of said antiligand or a compound conjugated to said antiligand which interacts with said label, and wherein a reagent is employed having both label and ligand or a ligand analog, where ligand analog is capable of specifically binding to said antiligand, said reagent comprising a liposome of amphiphilic molecules having from about 0.05 to 15 mole percent of ligand or ligand analog conjugated to an amphiphile and from 0.05 to 15 mole percent of a label conjugated to at least one of said amphiphile molecules; wherein said immunoassay comprises combining in an aqueous medium at a pH in the range of about 5 to 10 and at a temperature in the range of about 10° to 50° C.: a sample suspected of containing said analyte; said reagent; and antiligand, when ligand is the analyte or said antiligand is conjugated with a compound which interacts with said label; and determining the value of said detectible signal as compared to said value determined with a sample having a known amount of analyte.
Claim 10
Dependent Claim
19 words
A method according to claim 7, wherein said label is a fluorescer and antifluorescer is added to said medium.
Claim 11
Dependent Claim
13 words
A method according to claim 8, wherein said fluorescer is a fluorescein derivative.
Claim 12
Independent Claim
15 words
A method according to any of claims 7 to 9, wherein said ligand is cardiolipin.
Claim 13
Dependent Claim
19 words
A method according to claim 7, where from about 16 to 99.5 mole percent of said liposome is phospholipid.
Claim 14
Dependent Claim
16 words
A method according to claim 11, where up to 40 mole percent is a cholestanol derivative.
Claim 15
Dependent Claim
108 words
An immunoassay for the determination of the presence of anticardiolipin in human serum which comprises: combining in an aqueous buffered medium at a pH in the range of about 6.5 to 9.5: (1) a human serum sample suspected of containing anticardiolipin (2) liposomes comprised of from about 60 to 99.5 mole percent of a phosphatidyl compound, from about 0.05 to 5 mole percent of cardiolipin and from about 0.1 to 10 mole percent of a phosphatidyl-fluorescein conjugate; and (3) antifluorescein; and determining the amount of fluorescence from said assay medium as compared to the amount of fluorescence obtained from an assay medium having a known amount of anticardiolipin.
Claim 16
Dependent Claim
108 words
An immunoassay for the determination of the presence of anticardiolipin in human serum which comprises: combining in an aqueous buffered medium at a pH in the range of about 6.5 to 9.5: (1) a human serum sample suspected of containing anticardiolipin (2) liposomes comprised of from about 60 to 99.5 mole percent of a phosphatidyl compound, from about 0.05 to 5 mole percent of cardiolipin and from about 0.1 to 10 mole percent of a phosphatidyl-fluorescein conjugate; and (3) antifluorescein; and determining the amount of fluorescence from said assay medium as compared to the amount of fluorescence obtained from an assay medium having a known amount of anticardiolipin.
Claim 17
Dependent Claim
75 words
An immunoassay composition of matter comprising a liposome having from 16 to 99.5 mole percent of phospholipid, up to 40 mole percent of cholestanol, from about 0.05 to 15 mole percent of a fluorescent label bonded to a phospholipid, wherein said fluorescent label has an extinction coefficient of at least about 10 4 or at above 350 nm and a hapten of from about 125 to 2,000 molecular weight bonded to at least one phospholipid.
Claim 18
Dependent Claim
75 words
An immunoassay composition of matter comprising a liposome having from 16 to 99.5 mole percent of phospholipid, up to 40 mole percent of cholestanol, from about 0.05 to 15 mole percent of a fluorescent label bonded to a phospholipid, wherein said fluorescent label has an extinction coefficient of at least about 10 4 or at above 350 nm and a hapten of from about 125 to 2,000 molecular weight bonded to at least one phospholipid.
Claim 19
Dependent Claim
17 words
An immunoassay composition of matter according to claim 14, wherein said fluorescent label is a fluorescein derivative.
Claim 20
Dependent Claim
46 words
An immunoassay composition of matter having at least about 16 mole percent of phospholipid, up to 40 mole percent of a cholestanol, from 0.05 to 15 mole percent of a fluorescent label bonded to a phospholipid and from about 0.05 to 15 mole percent of cardiolipin.
Claim 21
Dependent Claim
46 words
An immunoassay composition of matter having at least about 16 mole percent of phospholipid, up to 40 mole percent of a cholestanol, from 0.05 to 15 mole percent of a fluorescent label bonded to a phospholipid and from about 0.05 to 15 mole percent of cardiolipin.
Claim 22
Dependent Claim
39 words
An immunoassay composition of matter comprising a liposome having from about 60 to 99.5 mole percent of a phospholipid, from about 0.05 to 5 mole percent of cardiolipin, and from about 0.1 to 10 mole percent of phosphatidyl-fluorescer conjugate.
Claim 23
Dependent Claim
39 words
An immunoassay composition of matter comprising a liposome having from about 60 to 99.5 mole percent of a phospholipid, from about 0.05 to 5 mole percent of cardiolipin, and from about 0.1 to 10 mole percent of phosphatidyl-fluorescer conjugate.