Claims
- 1. An assay method for the determination of an analyte that is a member of a specific binding pair (sbp member) in a sample suspected of containing said analyte, which method consists essentially of
- (a) combining in an assay medium
- (1) said sample
- (2) non-magnetic particles having bound thereto an sbp member that specifically binds to said analyte and to a labeled analyte or labeled analyte analog, such that said labeled analyte or labeled analyte analog can be separated from said assay medium in relation to the concentration of said analyte in said sample, and
- (3) magnetic particles having the stone charge as said non-magnetic particles, said magnetic particles being from about 10-250 nm in diameter;
- (b) adding to said
- (4) said labeled analyte or labeled analyte analog;
- (c) adding thereafter to said medium
- (5) a polyionic reagent for non-specifically coaggregating said non-magnetic particles and said magnetic particles to obtain coaggregated particles;
- (d) subjecting said assay medium to a magnetic field gradient to separate said coaggregated particles from said medium to obtain separated particles; and
- (e) examining said assay medium or said coaggregated particles for the presence or amount of said labeled analyte or labeled analyte analog, the presence or amount thereof indicating the presence or amount of said analyte in said sample.
- 2. The method of claim 1 wherein said non-magnetic particles are selected from the group consisting of biological particles and synthetic particles.
- 3. The method of claim 2 wherein said biological particles are cells.
- 4. The method of claim 3 wherein said cells are erythrocytes.
- 5. The method of claim 2 wherein said synthetic particles are selected from the group consisting of latex particles and liposomes.
- 6. The method of claim 1 wherein said magnetic particles have a diameter of 20 to 100 nm.
- 7. The method of claim 6 wherein said magnetic particles are coated with a serum protein prior to combination with assay medium.
- 8. The method of claim 1 wherein said magnetic particles are present in a ferrofluid prior to combination with said assay medium.
- 9. The method of claim 1 wherein said polyionic reagent is a polycation selected from a group consisting of polybrene, aminodextran, chitosan, and positively charged liposomes.
- 10. The method of claim 1 which further comprises the step of treating said separated particles with a releasing reagent to reverse their non-specific binding.
- 11. The method of claim 10 wherein said releasing reagent is a polyionic reagent selected from the group consisting of a polycationic or polyanionic reagents.
- 12. The method of claim 11 wherein said releasing reagent is polyacrylate, citrate, or a sulfate.
- 13. An assay method for the determination of an analyte that is a member of a specific binding pair (sbp member) and is a non-magnetic particle in a sample suspected of containing said analyte, which method consists essentially of the steps of
- (a) combining in an assay medium
- (1) said sample,
- (2) a labeled sbp member that specifically binds to said analyte, such that said labeled sbp member can be separated from said assay medium in relation to the concentration of said analyte in said sample, and
- (3) magnetic particles having the same charge as said non-magnetic particles, said magnetic particles being from about 10-250 nm in diameter;
- (b) adding to said assay medium
- (4) a polyionic reagent for non-specifically coaggregating said non-magnetic particles and said magnetic particles to obtain coaggregated particles;
- (c) subjecting said assay medium to a magnetic field gradient to separate said coaggregated particles from said medium to obtain separated particles; and
- (d) examining said assay medium or said coaggregated particles for the presence or amount of said labeled sbp member, the presence or amount thereof indicating the presence or amount of said analyte in said sample.
- 14. The method of claim 13 wherein said non-magnetic particles are biological particles.
- 15. The method of claim 14 wherein said biological particles are cells.
- 16. The method of claim 15 wherein said cells are erythrocytes.
- 17. The method of claim 13 wherein said magnetic particles have a diameter from about 20 to 100 nm.
- 18. The method of claim 17 wherein said magnetic particles are coated with a serum protein prior to combination with assay medium.
- 19. The method of claim 13 wherein said magnetic particles are present in a ferrofluid prior to combination with said assay medium.
- 20. The method of claim 13 wherein said polyionic reagent is a cation selected from a group consisting of polybrene, aminodextran, chitosan, and positively charged liposomes.
- 21. The method of claim 1 which further comprises the step of treating said separated particles with a releasing reagent to reverse their non-specific binding.
- 22. The method of claim 21 wherein said releasing reagent is a polyionic reagent selected from the group consisting of a polycationic or polyanionic reagents.
- 23. The method of claim 22 wherein said releasing reagent is polyacrylate, citrate, or a sulfate.
Parent Case Info
This is a is a continuation of application Ser. No. 07/787,847, filed Nov. 5, 1991, now abandoned, which is a division of application Ser. No. 07/533,622, filed Jun. 5, 1990, now U.S. Pat. No. 5,076,950, which in turn is a division of application Ser. No. 07/262,771, filed Oct. 26, 1988, now U.S. Pat. No. 4,935,147, which in turn is a continuation of application Ser. No. 06/811,202, filed Dec. 20, 1985, now abandoned.
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Divisions (3)
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Number |
Date |
Country |
Parent |
533622 |
Jun 1990 |
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Parent |
262771 |
Oct 1988 |
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Parent |
811202 |
Dec 1985 |
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Continuations (1)
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Number |
Date |
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Parent |
787847 |
Nov 1991 |
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