Claims
- 1. A method for determining cyclosporine, or cyclosporine and metabolites of cyclosporine, in a test sample, said method comprising the steps of:
- (a) contacting said test sample with a cyclosporine derivative corresponding to the formula: ##STR18## wherein ##STR19## represents a single or double bond; F1 is a detectable moiety from a luminescent molecule selected from the group consisting of fluoresceinamines and carboxyfluoresceins;
- X1 is a linking group of 1-15 atoms excluding hydrogen;
- R1 is hydrogen, OH or OCOR6; and
- R6 is an alkyl group of from 1-6 atoms or X1-F1; and an antibody capable of binding to (i) cyclosporine, or cyclosporine and metabolites of cyclosporine, and (ii) said cyclosporine derivative, to form a reaction solution therewith, said cyclosporine derivative capable of producing a detectable fluorescence polarization response to the presence of said antibody;
- (b) passing a plane of polarized light through the said reaction solution to obtain a fluorescence polarization response; and
- (c) detecting said fluorescent polarization response to said reaction solution as a function of the amount of cyclosporine, or cyclosporine and metabolites of cyclosporine, present in said test sample.
- 2. A method for determining cyclosporine, or cyclosporine and metabolites of cyclosporine, in a test sample, said method comprising the steps of:
- (a) contacting said test sample with a cyclosporine derivative corresponding to the formula: ##STR20## wherein ##STR21## represents a single or double bond; R7 is hydrogen or an acyl group of 1-6 carbon atoms;
- R8 is hydrogen or CH2OR7;
- X2 is a linking group of 1-30 atoms excluding hydrogen; and
- F1 is a detectable moiety from a luminescent molecule selected from the group consisting of fluoresceinamines and carboxyfluoresceins; and an antibody capable of binding to (i) cyclosporine, or cyclosporine and metabolites of cyclosporine, and (ii) said cyclosporine derivative, to form a reaction solution therewith, said cyclosporine derivative capable of producing a detectable fluorescence polarization response to the presence of said antibody;
- (b) passing a plane of polarized light through the said reaction solution to obtain a fluorescence polarization response; and
- (c) detecting said fluorescent polarization response to said reaction solution as a function of the amount of cyclosporine, or cyclosporine and metabolites of cyclosporine, present in said test sample.
- 3. A method for determining cyclosporine, or cyclosporine and metabolites of cyclosporine, in a test sample, said method comprising the steps of:
- (a) contacting said test sample with a cyclosporine derivative corresponding to the formula: ##STR22## wherein ##STR23## represents a single or double bond; R7 is hydrogen or an acyl group of 1-6 carbon atoms;
- R9 is hydrogen or OR7;
- X1 is a linking group of 1-15 atoms excluding hydrogen; and
- F1 is a detectable moiety from a luminescent molecule selected from the group consisting of fluoresceinamines and carboxyfluoresceins; and an antibody capable of binding to (i) cyclosporine, or cyclosporine and metabolites of cyclosporine, and (ii) said cyclosporine derivative, to form a reaction solution therewith, said cyclosporine derivative capable of producing a detectable fluorescence polarization response to the presence of said antibody;
- (b) passing a plane of polarized light through the said reaction solution to obtain a fluorescence polarization response; and
- (c) detecting said fluorescent polarization response to said reaction solution as a function of the amount of cyclosporine, or cyclosporine and metabolites of cyclosporine, present in said test sample.
- 4. A method for determining cyclosporine, or cyclosporine and metabolites of cyclosporine, in a test sample, said method comprising the steps of:
- (a) contacting said test sample with a cyclosporine derivative corresponding to the formula: ##STR24## wherein ##STR25## represents a single or double bond; R7 is hydrogen or an acyl group of 1-6 carbon atoms;
- X1 is a linking group of 1-15 carbon atoms excluding hydrogen; and
- F1 is a detectable moiety from a luminescent molecule selected from the group consisting of fluoresceinamines and carboxyfluoresceins; and an antibody capable of binding to (i) cyclosporine, or cyclosporine and metabolites of cyclosporine, and (ii) said cyclosporine derivative, to form a reaction solution therewith, said cyclosporine derivative capable of producing a detectable fluorescence polarization response to the presence of said antibody;
- (b) passing a plane of polarized light through the said reaction solution to obtain a fluorescence polarization response; and
- (c) detecting said fluorescent polarization response to said reaction solution as a function of the amount of cyclosporine, or cyclosporine and metabolites of cyclosporine, present in said test sample.
- 5. A method for determining cyclosporine, or cyclosporine and metabolites of cyclosporine, in a test sample, said method comprising the steps of:
- (a) contacting said test sample with a cyclosporine derivative corresponding to the formula: ##STR26## wherein ##STR27## represents a single or double bond; R7 is hydrogen or an acyl group of 1-6 carbon atoms;
- X1 is a linking group of 1-15 atoms excluding hydrogen; and
- F1 is a detectable moiety from a luminescent molecule selected from the group consisting of fluoresceinamines and carboxyfluorescein; and an antibody capable of binding to (i) cyclosporine, or cyclosporine and metabolites of cyclosporine, and (ii) said cyclosporine derivative, to form a reaction solution therewith, said cyclosporine derivative capable of producing a detectable fluorescence polarization response to the presence of said antibody;
- (b) passing a plane of polarized light through the said reaction solution to obtain a fluorescence polarization response; and
- (c) detecting said fluorescent polarization response to said reaction solution as a function of the amount of cyclosporine, or cyclosporine and metabolites of cyclosporine, present in said test sample.
- 6. A test kit useful for the fluorescent polarization immunoassay determination of the amount of cyclosporine, or cyclosporine and metabolites of cyclosporine, in a biological test sample, said test kit comprising:
- (a) a cyclosporine derivative corresponding to the formula: ##STR28## wherein ##STR29## represents a single or double bond; F1 is a detectable moiety from a luminescent molecule selected from the group consisting of fluoresceinamines and carboxyfluoresceins;
- X1 is a linking group of 1-15 atoms excluding hydrogen;
- R1 is hydrogen, OH or OCOR6; and
- R6 is an alkyl group of from 1-6 atoms or X1-F1;
- (b) an antibody capable of binding to (i) cyclosporine, or cyclosporine and metabolites of cyclosporine, and (ii) said cyclosporine derivative, said cyclosporine derivative capable of producing a detectable fluorescence polarization response to the presence of said antibody.
- 7. The test kit of claim 6 wherein said antibody is capable of binding to cyclosporine and said cyclosporine derivative.
- 8. The test kit of claim 6 wherein said antibody is capable of binding to cyclosporine and metabolites of cyclosporine and said cyclosporine derivative.
- 9. The test kit of claim 6, further comprising a precipitation reagent.
- 10. The test kit of claim 9, wherein said precipitation reagent comprises methanol, ethylene glycol and zinc sulfate.
- 11. The test kit of claim 6, further comprising a solubilization reagent for whole blood test samples.
- 12. The test kit of claim 11, wherein said solubilization reagent comprises alkyloxy(polyethyleneoxypropyleneoxy)-isopropanol.
- 13. The method of claim 1, wherein said antibody is capable of binding to cyclosporine and said cyclosporine derivative.
- 14. The method of claim 1, further comprising the step of treating said test sample with a precipitation reagent.
- 15. The method of claim 14, wherein said precipitation reagent comprises methanol, ethylene glycol and zinc sulfate.
- 16. The method of claim 1, wherein said test sample is whole blood.
- 17. The method of claim 16, further comprising the step of treating said whole blood test sample with a solubilization reagent.
- 18. The method of claim 17, wherein said solubilization reagent comprises alkyloxy(polyethyleneoxypropyleneoxy)-isopropanol.
Parent Case Info
This application is a Divisional of U.S. Ser. No. 08/148,164 filed Nov. 4, 1993, now U.S. Pat. No. 5,489,668, which is a Continuation of U.S. Ser. No. 07/952,488, filed Sep. 28, 1992 and now abandoned, which is a Continuation of U.S. Ser. No. 07/567,842, filed Aug. 15, 1990 and now abandoned.
US Referenced Citations (3)
Number |
Name |
Date |
Kind |
4727035 |
Mahoney |
Feb 1988 |
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5169773 |
Rosenthaler et al. |
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Wang et al. |
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Divisions (1)
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Number |
Date |
Country |
Parent |
148164 |
Nov 1993 |
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Continuations (2)
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Number |
Date |
Country |
Parent |
952488 |
Sep 1992 |
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Parent |
567842 |
Aug 1990 |
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