Fluorescent pigments for tagging biological molecules

• US 5,132,206 A
• Filed: 10/20/1988
• Issued: 07/21/1992
• Est. Priority Date: 06/11/1984
• Status: Expired due to Fees

• Current Assignee: William J. Dreyer
• Original Assignee: William J. Dreyer
• Inventors: Dreyer, William J.
• Primary Examiner(s): Yarbrough, Amelia Burgess
• Application Number:
  US07/262,184
• Time in Patent Office: 1,370 Days
• Field of Search: 435/6, 435/803, 435/7, 435/4, 436/94, 436/501, 436/518, 436/523, 436/532, 436/4, 436/800, 436/805, 436/809, 436/810, 436/818, 436/814, 422/61, 935/78, 935/81, 935/76
• US Class Current: 435/6.14
• CPC Class Codes: C12Q 1/6813 Hybridisation assays
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: Y10S 436/80 Fluorescent dyes, e.g. rhodamine
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10S 436/814 Pregnancy
• CPC Class Codes: Y10S 436/818 Human chorionic gonadotropin
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

A general method of assay for biological molecules using daylight fluorescent particles. The method described is applicable to assays involving immunological reagents, nucleic acids, hormones and neurotransmitters.

Claims

1. A method for assaying biological molecules comprising a support to which are bound on a portion of the surface of said support first molecules complementary to second molecules, said second molecules being said biological molecules, the balance of the surface of said support having bound thereto third molecules which are similar to the first molecules but not complementary to second molecules, the third molecules being attached to a portion of the support, and fourth molecules, also complementary to the second molecules, and labeled with a daylight fluorescent pigment, and causing the fourth molecules to become attached to those second molecules bound to first molecules.

2. The method of claim 1 wherein the assay is a pregnancy test, said second molecules are human chorionic gonadotropin, said fourth molecules cause daylight fluorescence only on said portion of the surface when human chorionic gonadotropin is present.

3. A method for assaying biological molecules according to claim 1 in which the substrate has a convoluted surface.

4. A method according to claim 1 in which the complementary molecule is an immunological ligand.

5. A method according to claim 1 in which the complementary molecule is a nucleic acid.

6. A method according to claim 1 in which the complementary molecule is one of a ligand receptor pair.

7. A method for assaying of biological molecules in which first molecules, complementary to the biological molecules which are to be assayed, are bound to a surface dyed with a first color, the method comprising the steps of:

(a) exposing the surface coated with the first molecule to an unknown solution thought to contain second molecules, said second molecules being biological molecules which are to be assayed.

(b) adding to this solution daylight fluorescent pigmented microparticles which have been coated with third molecules also complementary to the second molecules which are to be assayed; and

(c) comparing the resulting color with a predetermined list of colors and corresponding concentrations of the second molecules, to obtain an indication of the concentration of the second molecules.

8. A method according to claim 7 in which the labeled molecules are immunological ligands.

9. A method according to claim 7 in which the labeled molecules are nucleic acids.

10. A method according to claim 7 in which each of the labeled molecules is a receptor of the first molecules.

11. A method according to claim 7 in which the substrate to which the first molecules are bound have a convoluted surface.

12. A method for assaying of biological molecules in which a plurality of first molecules complementary to a second molecule to be assayed are bound to the surface of a convoluted support in a container in which a plurality of third molecules which are the second molecules or analogues thereof are bound to the first molecules, said second molecules being biological molecules, and in which the third molecules are covalently bound to the surfaces of daylight fluorescent pigmented microparticles, the method comprising the steps of:

(a) exposing first and third molecules to an aqueous solution containing second molecules to be assayed,

(b) allowing second molecules to bind to the first molecules; and

(c) determining the number of third molecules released from said surface.

13. A method according to claim 12 in which the first, the second, and the third molecules are immunological ligands.

14. A method according to claim 12 in which the first, the second, and the third molecules are nucleic acids.

15. A method for assaying first biological molecules comprising labeling second molecules which are complementary to the first biological molecules with microparticles exhibiting daylight fluorescence, and causing the labeled second molecules to contact the first biological molecules.

16. A method of labeling cells by contacting cells with molecules bound to microparticles exhibiting daylight fluorescence.

17. A method for assaying of biological molecules in which first molecules, complementary to the biological molecules which are to be assayed, are bound to a dipstick comprising a substrate containing a plurality of discrete areas therein, each of said areas having bonded to the surface thereof said first molecules, said method comprising:

(a) contacting said dipstick into a specimen thought to contain said biological molecules,

(b) contacting the dipstick with third molecules labeled with a daylight fluorescent pigment, said third molecule being complementary to the biological molecules, and

(c) examining the dipstick for presence of daylight fluorescence in any of said areas.