Hapten derivatized capture membrane and diagnostic assays using such membrane

• US 5,888,728 A
• Filed: 06/07/1995
• Issued: 03/30/1999
• Est. Priority Date: 10/17/1988
• Status: Expired due to Term

• Current Assignee: Molecular Devices
• Original Assignee: Molecular Devices
• Inventors: Zuk, Robert F., Sheldon, Edward L., Kung, Viola T., Burke, Charles R., Olson, John D., Armenta, Richard D.
• Primary Examiner(s): Marschel, Ardin H.
• Assistant Examiner(s): Riley, Jezia
• Application Number:
  US08/484,968
• Time in Patent Office: 1,392 Days
• Field of Search: 435/6, 435/172.3, 436/501, 436/530, 436/532, 436/543, 436/63, 436/823, 935/77, 935/78, 935/110, 536/22.1, 536/25.32
• US Class Current: 435/6.14
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07K 14/765 Serum albumin, e.g. HSA
• CPC Class Codes: C12Q 1/6834 Enzymatic or biochemical coupling of nucleic acids to a solid phase
• CPC Class Codes: C12Q 2563/131 the label being a member of a cognate binding pair, i.e. extends to antibodies, haptens, avidin
• CPC Class Codes: G01N 33/5308 for analytes not provided for elsewhere, e.g. nucleic acids, uric acid, worms, mites
• CPC Class Codes: G01N 33/537 with separation of immune complex from unbound antigen or antibody
• CPC Class Codes: G01N 33/54306 Solid-phase reaction mechanisms
• CPC Class Codes: G01N 33/54353 with ligand attached to the carrier via a chemical coupling agent coatings G01N33/54393
• CPC Class Codes: Y10S 436/823 Immunogenic carrier or carrier per se

Abstract

The present invention encompasses a capture membrane comprising a porous filter membrane having a hapten bound directly or indirectly to the membrane wherein complexes formed by specific binding having an anti-hapten bound to a binding member of the specifically binding complex are removed from a solution by the hapten as the solution passes through the membrane. In the preferred embodiment biotin is the hapten and avidin or streptavidin is the anti-hapten.

Claims

1. A method for determining single or double stranded DNA in a mixture suspected of containing said DNA, comprising:

(a) preparing a solution of first and second binding members and a portion of the mixture suspected to contain said DNA,

the first binding member being anti-DNA, single-stranded binding protein, or an oligonucleotide probe complimentary to at least a portion of the DNA directly or indirectly bound to avidin or streptavidin,

the second binding member being anti-DNA, single-stranded binding protein, or an oligonucleotide probe complimentary to at least a portion of the DNA directly or indirectly bound to a detectable label,

where the binding members in the presence of DNA in the mixture form a specifically binding complex between the DNA and the binding members;

(b) filtering the solution through a filter membrane having adhered thereto a compound of the formula;

##STR2## where n is an integer of at least 5; and

(c) detecting the label.

3. A method according to claim 1, wherein n is an integer of from 5 to about 20.

4. A method according to claim 1, wherein the protein is Bovine Serum Albumin.

5. A method according to claim 4, wherein n is from 5-20.

6. A methods according to claim 3, wherein the protein is Bovine Serum Albumin.

7. A method according to claim 1, wherein the binding members are antibodies.

8. A method according to claim 1, wherein the membrane comprises nitrocelluloses.

9. A method according to claim 1, wherein the label is an enzyme.

10. A method according to claim 1, wherein the label is detected by an electrode.

11. A method according to claim 10, wherein the electrode is a semiconductor.

20. A method according to claim 1, wherein the first binding memberis biotin-anti-DNA.

21. A method according to claim 1, wherein the second binding member is labeled single stranded binding protein.

22. A method according to claim 1, wherein the second binding member is labeled anti-DNA.

23. A method according to claim 1, wherein the first binding member is biotin-single stranded binding protein.

2. A method for determining single or double stranded DNA in a mixture suspected of containing said DNA, comprising:

(a) preparing a solution of a binding member linked to avidin or streptavidin, the binding member having a binding affinity for the DNA to be determined; and

(b) filtering the solution through a filter membrane having adhered thereto a compound of the formula;

##STR3## where n is an integer of at least 5.

12. A method according to claim 2, further comprising (c) filtering a solution suspected to contain the substance to be determined through the membrane.

13. A method according to claim 12, further comprising (d) filtering a solution of a detectable label linked to a binding member, the binding member having a binding affinity for the substance to be determined.

14. A method according to claim 13, further comprising detecting the label .

15. A method according to claim 2, wherein the protein is Bovine Serum Albumin.

16. A method according to claim 15, wherein n is from 5-20.

17. A method according to claim 2, further comprising (c) filtering a second solution through the membrane, the second solution containing a detectable label linked to a binding member, the binding member having a binding affinity for the substance to be determined, and the second solution being suspected to contain the substance to be determined.

18. A membrane according to claim 17, wherein the porous membrane comprises nitrocellulose.

19. A method according to claim 13, wherein the porous membrane comprises nitrocellulose.

24. A method according to claim 2, wherein the first binding member is biotin-anti-DNA.

25. A method according to claim 2, wherein the second binding member is labeled single stranded binding protein.

26. A method according to claim 2, wherein the second binding member is labeled anti-DNA.

27. A method according to claim 2, wherein the first binding member is biotin-single stranded binding protein.