Solid phase assay

• US 4,743,560 A
• Filed: 03/26/1984
• Issued: 05/10/1988
• Est. Priority Date: 03/26/1984
• Status: Expired due to Fees

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Campbell, Robert L., O'Connell, James P., Wagner, Daniel B.
• Primary Examiner(s): Marantz, Sidney
• Assistant Examiner(s): Saunders, David A.
• Application Number:
  US06/593,411
• Time in Patent Office: 1,506 Days
• Field of Search: 436/501, 436/518, 436/519, 436/520, 436/523, 436/528, 436/532, 436/535, 436/829, 436/530, 436/534, 436/510, 436/800, 436/808, 436/815, 436/818, 435/7, 435/177, 435/180, 435/182, 435/810
• US Class Current: 436/501
• CPC Class Codes: G01N 33/48 Biological material, e.g. blood, urine G01N33/02, G01N33/26, G01N33/44, G01N33/46 take precedence; Haemocytometers counting blood corpuscules distributed over a surface by scanning the surface G06M11/02
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: G01N 33/586 Liposomes, microcapsules or cells
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/815 Test for named compound or class of compounds
• CPC Class Codes: Y10S 436/818 Human chorionic gonadotropin
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation

Abstract

Solid phase assay for an analyte wherein binder is supported on a solid support, such as nitrocellulose, and the tracer is comprised of ligand labeled with a particulate label, such as a liposome, including a detectable marker which is not visible.

Claims

1. A process for assaying for an analyte, comprising:

contacting a binder supported on a test area of a solid support with analyte and a tracer, said binder being a specific binder for at least the analyte, said tracer being comprised of a ligand labeled with a liposome, said liposome including a detectable marker which is not visible, said ligand being bound to one of the binder on the support or the analyte bound to the binder on the support, said test area being formed of a material having a surface area for supporting the binder and said binder being supported in the test area in a concentration of at least 1 ug/cm and at which tracer may be determined without lysis of the liposome; and

determining the tracer bound in said test area as a measure of analyte in a sample.

2. The process of claim 1 wherein analyte and tracer are sequentially contacted with the binder on the solid support.

3. The process of claim 1 wherein the analyte and tracer are simultaneously contacted with binder on the solid support.

4. The process of claim 1 wherein the analyte is digoxin.

5. The process of claim 1 wherein the analyte is (hCG) human chorionic gonadotropin.

6. The process of claim 1 wherein the tracer is determined without lysis of the liposome.

7. The process of claim 1 wherein the solid support is in sheet form.

8. The process of claim 1 wherein the binder is an antibody.

9. The process of claim 8 wherein the tracer is a labeled form of the analyte.

10. The process of claim 8 wherein the tracer is a labeled form of an antibody for the analyte.

11. The process of claim 1 wherein the assay is for human chorionic gonadotropin the binder is antibody to human chorionic gonadotropin and said tracer is comprised of antibody to human chorionic gonadotropin labeled with a (sac) liposome containing a fluorescent material as said detectable marker.

12. The process of claim 11 wherein the test area is formed of nitrocellulose.

13. The process of claim 1 wherein the assay is for digoxin, said binder is antibody to digoxin and the tracer is comprised of an analogue of digoxin (bound by said antibody) labeled with a sac containing a fluorescent material as said detectable marker(.), said analogue being bound by said antibody.

14. The process of claim 13 wherein the test area is formed of nitrocellulose.

15. A process for assaying for an analyte, comprising:

contacting a binder supported on a test area of a solid support with analyte and a tracer, said binder being a binder specific for at least the analyte, said tracer being comprised of a ligand labled with a liposome including a detectable marker which is not visible, said ligand being bound to one of the binder on the support or the analyte bound to the binder on the support, said test area being formed of nitrocellulose, said binder being supported in the test area in a concentration of at least 1 ug/cm and at which tracer may be determined without lysis of the liposome; and

determining the tracer bound in said test area as a measure of analyte in a sample.

16. The process of claim 15 wherein the tracer in said test area is determined without lysis of the liposome.

17. The process of claim 16 wherein the analyte is (hCG) human chorionic gonadotropin.

18. The process of claim 16 wherein the analyte is digoxin.

19. The process of claim 16 wherein the analyte is in a sample in a concentration of no greater than 10^-9 gm/ml

20. The process of claim 15 wherein the analyte is in a sample in a concentration of no greater than 10^-9 gm/ml.

21. The process of claim 15 wherein the solid support is in sheet form.

22. The process of claim 15 wherein the support contains a plurality of said test areas.

23. The process of claim 15 wherein the binder is supported in a concentration of at least 40 μ

g/cm².

24. The process of claim 23 wherein the binder is in the test area as a spot having a diameter of from 3 to 5 mm.

25. A reagent kit for determining an analyte comprising:

a solid support including a binder on a test area of a solid support; and

a tracer, said binder being a binder specific for at least the analyte, said tracer being comprised of a ligand labeled with a liposome containing a detectable marker which is not visible, said ligand being bound to one of the binder on the support or the analyte bound to the binder on the support during an assay, said test area being formed of a material having a surface area supporting the binder and the binder being supported in a concentration of at least 1 ug/cm and at which tracer may be determined without lysis of the liposome.

26. The kit of claim 25 wherein the (binder) test area is formed of nitrocellulose.

27. The kit of claim 26 wherein the label of the tracer is a liposome including a fluorescent material as said marker.

28. The kit of claim 26 wherein the binder is an antibody.

29. The kit of claim 28 wherein the analyte is digoxin, the binder is antibody to digoxin and the ligand of the tracer is an analogue of digoxin.

30. The kit of claim 28 wherein the analyte is (hCG) human chorionic gonadotropin.

31. The kit of claim 30 wherein the binder is antibody to (hCG) human chorionic gonadotropin and the ligand of the tracer is antibody to (hCG) human chorionic gonadotropin.

32. A process for assaying for an analyte, comprising:

contacting a binder supported on a test area of a solid support with analyte and a tracer, said binder being a specific binder for at least the analyte, said tracer being comprised of a ligand labeled with a lipsome, said liposome including a detectable marker which is not visible, said test area being formed of a material having a surface area for supporting the binder and the binder being supported in the test area in a concentration of at least 1 ug/cm and at which tracer may be determined without lysis of the liposome; and

determining the tracer bound in said test area as a measure of analyte in a sample.

33. The process of claim 32 wherein the test area is formed of nitrocellulose.

34. The process of claim 32, wherein the tracer is determined without lysis of the liposome.