Claims
- 1. An apparatus for determining at least one analyte in a fluid sample, comprising:
- (a) a first porous zone comprising an immobilized, non diffusible reagent capable of specifically binding to at least one diffusible labeled reactant comprising a label conjugated to either said analyte, an analogue of said analyte or a specific binding partner for said analyte, and
- (b) a second porous zone comprising (i) a diffusible interactive reagent capable of reacting with the label portion of said diffusible labeled reactant to produce a diffusible detectable moiety and (ii) a means for separating said diffusible interactive reagent for said diffusible detectable moiety.
- 2. The apparatus of claim 1, wherein said first and second zones are in at least partial fluid contact with each other.
- 3. The apparatus of claim 1 wherein said first porous zone comprises two parts,
- (a') a first part comprising said at least one diffusible labeled reactant and
- (a'') a second part comprising said immobilized, non-diffusible reagent which specifically binds to said at least one diffusible labeled reactant.
- 4. The apparatus of claim 1, wherein said second porous zone comprises two parts,
- (b') a first part comprising said reactive component and
- (b'') a second part comprising said separating means.
- 5. The apparatus of claim 1, wherein said first and second porous zones each comprise two parts,
- (a') a first part of said first porous zone comprising a labeled, diffusible, specific binding partner for said analyte and
- (a'') a second part of said first porous zone comprising said immobilized, non-diffusible reagent wherein said immobilized, non-diffusible reagent is capable of specifically binding to any labeled binding partner unbound to analyte,
- (b') a first part of said second porous zone comprising said diffusible interactive reagent capable of reacting with the label portion of said diffusible labeled reactant to produce a detectable moiety, and
- (b'') a second part of said second porous zone comprising a means for separating said diffusible interactive reagent from said diffusible detectable moiety.
- 6. The apparatus of claim 3, wherein the first and second parts of said first porous zone are in at least partial fluid contact with each other.
- 7. The apparatus of claim 4, wherein the first and second parts of said second porous zone are in at least partial fluid contact with each other.
- 8. The apparatus of claim 5, wherein the second part of said first porous zone and the first part of said second porous zone are in at least partial fluid contact with each other.
- 9. The apparatus of claim 5, wherein the first part of said first porous zone is in at least partial fluid contact with the second part of said first porous zone, the second part of said first porous zone is in partial fluid contact with the first part of said second porous zone, and the second part of said second porous zone is in partial fluid contact with the first part of said second porous zone.
- 10. The apparatus of claim 1 wherein said separating means is an ion exchange paper capable of selectively retaining either said diffusible interactive reagent or said diffusible detectable moiety.
- 11. The apparatus of claim 1 wherein said first zone further comprises said at least one diffusible, labeled specific binding partner for said analyte.
- 12. The apparatus of claim 1 wherein said first zone further comprises either said at least one diffusible labeled analyte or said at least one diffusible labeled analogue of said analyte.
- 13. The apparatus of claim 1, wherein said first zone further comprises a plurality of different diffusible, labeled specific binding partners each type of which is capable of specifically binding to a separate analyte to be determined.
- 14. The apparatus of claim 1, wherein said first porous zone further comprises a plurality of one of (i) diffusible labeled analytes or (ii) diffusible labeled analyte analogues.
- 15. The apparatus of claim 11 wherein said at least one diffusible labeled specific binding partner for said analyte comprises an antibody or binding fragment thereof.
- 16. The apparatus of claim 13, wherein each of said diffusible, labeled specific binding partners comprises an antibody or binding fragment thereof.
- 17. The apparatus of claim 13, wherein said second porous zone further comprises a plurality of different diffusible interactive reagents, each type of which is capable of reacting with a specific diffusible, labeled binding partner to produce a specific diffusible, detectable moiety.
- 18. The apparatus of claim 14, wherein said second porous zone further comprises a plurality of diffusible, interactive reagents, each of which is capable of reacting with the label portion of a specific diffusible labelled analyte or a specific diffusible labeled analogue of said analyte.
- 19. An apparatus for determining at least one analyte in a fluid sample, comprising:
- (a) a first zone comprising an immobilized, non-diffusible reagent capable of specifically binding to at least one diffusible reactant comprising analyte, labeled analyte, labeled analyte analogue or labeled analyte reaction partner,
- (b) a second zone comprising (i) a diffusible interactive reagent capable of reacting with the label of said diffusible detectable moiety and (ii) a means for separating said diffusible interactive reagent from said diffusible detectable moiety, and
- (c) at least one flow regulating means which regulates movement of a fluid sample introduced to said apparatus, wherein said flow regulating means has a flow rate different from the flow rate of at least a part of one of said first and second zones.
- 20. The apparatus of claim 19, wherein said first and second porous zones are in at least partial fluid contact with each other.
- 21. The apparatus of claim 19, wherein said first zone comprises two parts,
- (a') a first part comprising said at least one diffusible labeled reactant, and
- (a'') a second part comprising said immobilized, non-diffusible reagent which specifically binds to said at least one diffusible labeled reactant.
- 22. The apparatus of claim 19, wherein said second porous zone comprises two parts,
- (b') a first part which comprises said diffusible interactive reagent capable of reacting with the labeled portion of said diffusible labeled reactant, and
- (b'') a second part which comprises said flow regulating means.
- 23. The apparatus of claim 19, wherein each of said first and second porous zones comprises two parts, wherein said first zone comprises:
- (a') a first part comprising said at least one diffusible labeled reactant, and
- (a'') a second part comprising said immobilized, non-diffusible reagent which specifically binds to said at least one diffusible labeled reactant, and said second zone comprises:
- (b') a first part which comprises said diffusible interactive reagent capable of reacting with the labeled portion of said diffusible labeled reactant, and
- (b'') a second part which comprises said flow regulating means.
- 24. The apparatus of claim 19, wherein said flow regulating meas is incorporated in said first porous zone.
- 25. The apparatus of claim 19, wherein said flow regulating means is incorporated in said second porous zone.
- 26. The apparatus of claim 19, wherein said flow regulating means is positioned between said first and second porous zones.
- 27. The apparatus of claim 19, wherein said flow regulating means is positioned in said apparatus before said first porous zone and said second porous zone is positioned in said apparatus after said first zone.
- 28. The apparatus of claim 21, wherein the first and second parts of said first porous zone are in at least partial fluid contact with each other.
- 29. The apparatus of claim 22, wherein the first and second parts of said second porous zone are in at least partial fluid contact with each other.
- 30. The apparatus of claim 23, wherein the second part of said first porous zone and the first part of said second porous zone are in at least partial fluid contact with each other.
- 31. The apparatus of claim 23, wherein the first part of said first porous zone is in at least partial fluid contact with the second part of said first porous zone, said second part of said first porous zone is in partial fluid contact with the first part of said second porous zone, and said second part of said second porous zone is in partial fluid contact with the first part of said second porous zone.
- 32. The apparatus of claim 19, wherein said flow regulating means comprises filter paper.
- 33. The apparatus of claim 19, wherein said first zone further comprises at least one diffusible, labeled reactant comprising a label conjugated to a specific binding partner for said analyte.
- 34. The apparatus of claim 19, wherein said first zone further comprises at least one diffusible, labeled reactant comprising a label conjugated to either of said analyte or an analogue of said analyte.
- 35. The apparatus of claim 19, wherein said first zone comprises a plurality of diffusible labeled reactants, each type of which comprises a distinct label conjugated to a specific binding partner for a particular analyte.
- 36. The apparatus of claim 19, wherein said first zone further comprises a plurality of distinct types of diffusible labeled reactants, each type of which comprises a label conjugated to a different analyte or analyte analogue.
- 37. The apparatus of claim 33, wherein said at least one diffusible labeled specific binding partner for said analyte comprises an antibody or bindable fragment thereof.
- 38. The apparatus of claim 35, wherein said plurality of diffusible labeled specific binding partners for said analyte comprises antibodies or binding fragments thereof.
- 39. The apparatus of claim 35, wherein said second zone comprises a plurality of multiple, distinct diffusible interactive reagents, each of which is capable of reacting with a distinct label of said plurality of diffusible labeled reacts to produce a distinct diffusible detectable moiety.
- 40. The apparatus of claim 36, wherein said second zone comprises a plurality of multiple, distinct diffusible interactive reagents, each of which is capable of reacting with a distinct label of said plurality of labeled reactants.
- 41. The apparatus of claim 31, wherein said flow regulating means is positioned in said apparatus before said first process zone and said second porous zone is positioned after said first porous zone.
- 42. Method for determining at least one analyte in a fluid sample, comprising:
- (a) contacting said sample with a labeled binding partner which specifically binds to said analyte to form a mixture comprising (i) complexes of analyte and labeled binding partner and (ii) uncomplexed labeled binding partner,
- (b) contacting said mixture to an immobilized reagent which binds to uncomplexed labeled binding partner but not complexes of analyte and labeled binding partner, so as to remove uncomplexed labeled binding partner from said mixture,
- (c) contacting said mixture to an apparatus comprising a diffusible, interactive reagent capable of reacting with the label portion of said labeled binding partner to produce a detectable moiety in said mixture,
- (d) contacting said mixture with a means for separating detectable moiety from any unreacted diffusible interactive reagent, and
- (e) measuring one of unreacted diffusible interactive reagent, removed uncomplexed labeled binding partner or detectable moiety as a measure of analyte in said sample.
- 43. Method for determining at least one analyte in a fluid sample, comprising:
- (a) contacting said sample with a labeled reactant comprising a label conjugated to either said analyte or an analogue of said analyte, to form a mixture;
- (b) contacting said mixture with an immobilized binding partner capable of binding to said analyte and to said labeled reagent to form complexes of (i) immobilized binding partner and analyte, and of (ii) immobilized binding partner and labeled reagent,
- (c) contacting said complex containing mixture with an interactive reagent capable of reacting with the label portion of said labeled reactant to form a detectable moiety,
- (d) contacting said complex containing mixture with means for separating unreacted interactive reagent and detectable moiety from each other, and
- (e) measuring one of unreacted interactive reagent, immobilized labeled reagent or detectable moiety as a measure of analyte in said sample.
- 44. Method for determining at least one analyte in a fluid sample, comprising:
- (a) contacting said sample with a labeled reactant comprising a label conjugated to either said analyte or an analogue of said analyte, to form a mixture;
- (b) contacting said mixture with an immobilized binding partner capable of binding to said analyte and to said labeled reagent, to form complexes of immobilized binding partner and analyte, and of immobilized binding partner and labeled reagent;
- (c) separating said complexes from said mixture;
- (d) contacting complex free mixture with an interactive reagent capable of reacting with the label portion of said labeled reactant to produce a detectable moiety;
- (e) contacting said complex free mixture with a means for separating unreacted interactive reagent from said detectable moiety, and
- (f) measuring one of label in said separated complex detectable moiety or unreacted interactive reagent as a measure of analyte in said sample.
- 45. Method for determining at least one analyte in a fluid sample, comprising:
- (a) contacting said sample with a complex of labeled reactant capable of specifically binding to said analyte and an immobilized binding partner for said labeled reactant, so as to form a mixture of displaced labeled reactant complexed to said analyte and free labeled reactant;
- (b) contacting said mixture to an immobilized reagent capable of specifically binding to free labeled reactant but not to complexes of labeled reactant and analyte, so as to remove free labeled reactant therefrom;
- (c) contacting said mixture to an interactive reagent which reacts with the label of said labeled reactant to form a detectable moiety;
- (d) contacting said mixture with a separation means for separating interactive reagent from detectable moiety; and
- (e) measuring immobilized labeled reactant, unreacted interactive reagent or detectable moiety as a measure of analyte in said sample.
Parent Case Info
This application is a continuation, of application Ser. No. 146,345, filed Jan. 21, 1988, now abandoned.
US Referenced Citations (6)
Foreign Referenced Citations (1)
Number |
Date |
Country |
8702774 |
Jul 1987 |
WOX |
Continuations (1)
|
Number |
Date |
Country |
Parent |
146345 |
Jan 1988 |
|