Heterologous system for the detection of chemically labeled DNA and other biological materials providing a receptor or target moiety thereon

• US 4,889,798 A
• Filed: 02/13/1987
• Issued: 12/26/1989
• Est. Priority Date: 02/09/1984
• Status: Expired due to Term

• Current Assignee: Enzo Diagnostics, Inc.
• Original Assignee: Enzo Biochem Incorporated
• Inventors: Rabbani, Elazar
• Primary Examiner(s): Brown, Johnnie R.
• Assistant Examiner(s): NOT, DEFINED
• Application Number:
  US07/015,563
• Time in Patent Office: 1,047 Days
• Field of Search: 435/6, 435/7, 435/188, 435/810, 435/803, 436/501, 436/518, 436/808, 436/824, 436/827, 935/78, 536/27, 530/367, 530/395, 530/396, 548/303
• US Class Current: 435/6.16
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: G01N 33/535 with enzyme label or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates
• CPC Class Codes: Y10S 435/803 Physical recovery methods, e.g. chromatography, grinding
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/824 Immunological separation techniques
• CPC Class Codes: Y10S 436/827 Lectins

Abstract

In accordance with the practices of this invention, there is provided a heterologous detection system and components useful in conjunction therewith and kits for carrying out the heterologous detection system. The heterologous detection system employs a heterologous entity and a signalling entity. The heterologous entity contains two free complex forming sites that form different complexes, i.e., two kinds of complexes can be formed. The first free complex forming site is utilized to recognize the labeled probe and the second free complex forming site is utilized to recognize the signalling entity, with each complex formed being different. The signalling entity contains a free complex forming site that can recognize the second free complex forming site of the heterologous entity and a signalling portion that is capable of generating a signal. An example of a heterologous detection system that can be utilized to detect biotinylated probe is a system that contains a heterologous entity comprising avidin and a lectin and a signalling entity comprising a glycosylated enzyme. The avidin can recognize the biotin of the probe and the lectin can recognize the glycosyl group of the enzyme. The enzyme is capable of generating a signal when treated with a suitable reagent. Such a heterologous detection system provides both a quick and highly sensitive technique for detecting a biotinylated probe.

Claims

1. A composition for detecting the presence or absence of an analyte in a sample, which composition comprises:

a biotinylated polynucleotide specifically bindable with said analyte;

a heterologous entity comprising streptavidin and a lectin; and

a signaling entity which comprises a third free complex forming site which is specifically bindable with said lectin and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

4. The composition of claims 1, 2 or 3 wherein said lectin is selected from the group consisting of Concanavalin A, soybean lectin, wheatgerm lectin, lotus seed lectin, potato lectin, dilichos biflorus agglutinin and lentil lectin.

5. The composition of claim 4 wherein said lectin is Concanavalin A.

2. A composition for detecting the presence or absence of an analyte in a sample, which composition comprises:

a biotinylated polynucleotide specifically bindable with said analyte;

a heterologous entity comprising avidin and a lectin; and

a signaling entity which comprises a third free complex forming site which is specifically bindable with said lectin and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

3. A composition for detecting the presence or absence of an analyte in a sample, which composition comprises:

a biotinylated polynucleotide specifically bindable with said analyte;

a heterologous entity comprising a lectin that is complexed with a glycosylated enzyme that is biotinylated;

or streptavidin; and

a signaling entity which comprises a third free complex forming site which is specifically bindable with said glycosylated enzyme and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

6. A composition for detecting the presence or absence of an analyte in a sample, which composition comprises:

a biotinylated polynucleotide specifically bindable with said analyte;

a heterologous entity comprising avidin or streptavidin complexed with a biotinylated enzyme that is glycosylated; and

a signaling entity which comprises a third free complex forming site which is specifically bindable with said glycosylated enzyme and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

7. A composition for detecting the presence or absence of an analyte in a sample, which composition comprises:

a biotinylated polynucleotide specifically bindable with said analyte;

a heterologous entity comprising streptavidin and a glycosyl group; and

a signaling entity which comprises a third free complex forming site which is specifically bindable with said glycosyl group and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

8. A composition for detecting the presence or absence of an analyte in a sample, which composition comprises:

a glycosylated polynucleotide specifically bindable with said analyte;

a heterologous entity comprising avidin and a lectin; and

a signaling entity which comprises a third free complex forming site which is specifically bindable with said avidin and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

9. A composition for detecting the presence or absence of an analyte in a sample, which composition comprises:

a glycosylated polynucleotide specifically bindable with said analyte;

a heterologous entity comprising streptavidin and a lectin; and

a signaling entity which comprises a third free complex forming site which is specifically bindable with said streptavidin and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

10. A composition for detecting the presence or absence of an analyte in a sample, which composition comprises:

a glycosylated polynucleotide specifically bindable with said analyte;

a heterologous entity comprising biotin and a lectin; and

a signaling entity which comprises a third free complex forming site which is specifically bindable with said biotin and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

11. The composition of claims 8, 9, or 10 wherein said lectin is selected from the group consisting of Concanavalin A, soybean lectin, wheatgerm lectin, lotus seed lectin, potato lectin, dilicho biflorus agglutinin and lentil lectin.

12. The composition of claim 11 wherein said lectin is Concanavalin A.

13. A kit useful for detecting the presence or absence of an analyte in a sample, which kit comprises, in packaged combination, the following:

(a) a first container having therein a labeled probe which comprises a biotinylated polynucleotide;

(b) a second container having therein a heterologous entity which comprises streptavidin and a lectin;

(c) a third container having therein a signalling entity which comprises a third free complex forming site which is specifically bindable with said lectin and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

16. The kit of claims 13, 14 or 15 wherein said lectin is selected from the group consisting of Concanavalin A, soybean lectin, wheatgerm lectin, lotus seed lectin, potato lectin, dilichos biflorus agglutinin and lentil lectin.

17. The kit of claim 16 wherein said lectin is Concanavalin A.

14. A kit useful for detecting the presence or absence of an analyte in a sample, which kit comprises, in packaged combination, the following:

(a) a first container having therein a labeled probe which comprises a biotinylated polynucleotide;

(b) a second container having therein a heterologous entity which comprises avidin and a lectin;

(c) a third container having therein a signalling entity which comprises a third free complex forming site which is specifically bindable with said lectin and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

15. A kit useful for detecting the presence or absence of an analyte in a sample, which kit comprises, in packaged combination, the following:

(a) a first container having therein a labeled probe which comprises a biotinylated polynucleotide;

(b) a second container having therein a heterologous entity which comprises a lectin that is complexed with a glycosylated enzyme that is biotinylated;

a third container having therein a signalling entity which comprises a third free complex forming site which is specifically bindable with said lectin and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

18. A kit useful for detecting the presence or absence of an analyte in a sample, which kit comprises, in packaged combination, the following:

(a) a first container having therein a labeled probe which comprises a biotinylated polynucleotide;

(b) a second container having therein a heterologous entity which comprises streptavidin and a glycosyl group;

(c) a third container having therein a signalling entity which comprises a third free complex forming site which is specifically bindable with said glycosyl group and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

19. A kit useful for detecting the presence or absence of an analyte in a sample, which kit comprises, in packaged combination, the following:

(a) a first container having therein a labeled probe which comprises a biotinylated polynucleotide;

(b) a second container having therein a heterologous entity which comprises avidin or streptavidin complexed with a biotinylated enzyme that is glycosylated;

(c) a third container having therein a signalling entity which comprises a third free complex forming site which is specifically bindable with said biotinylated enzyme that is glycosylated and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

20. A kit useful for detecting the presence or absence of an analyte in a sample, which kit comprises, in packaged combination, the following:

(a) a first container having therein a labeled probe which comprises a glycosylated polynucleotide;

(b) a second container having therein a heterologous entity which comprises avidin and a lectin;

(c) a third container having therein a signalling entity which comprises a third free complex forming site which is specifically bindable with said avidin and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

23. The kit of claims 20, 21 or 22 wherein said lectin is selected from the group consisting Concanavalin A, soybean lectin, wheatgerm lectin, lotus seed lectin, potato lectin, dilichos bilforus agglutinin and lentil lectin.

24. The kit of claim 23 wherein said lectin is Concanavalin A.

21. A kit useful for detecting the presence or absence of an analyte in a sample, which kit comprises, in packaged combination, the following:

(a) a first container having therein a labeled probe which comprises a glycosylated polynucleotide;

(b) a second container having therein a heterologous entity which comprises streptavidin and a lectin;

(c) a third container having therein a signalling entity which comprises a third free complex forming site which is specifically bindable with said streptavidin and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.

22. A kit useful for detecting the presence or absence of an analyte in a sample, which kit comprises, in packaged combination, the following:

(a) a first container having therein a labeled probe which comprises a glycosylated polynucleotide;

(b) a second container having therein a heterologous entity which comprises biotin and a lectin;

(c) a third container having therein a signalling entity which comprises a third free complex forming site which is specifically bindable with said biotin and, attached thereto, a signal generating portion capable of forming a signal associated with detection of said analyte.