Granted Patent B2

US 8,420,400 · App. 11/954,055 · Granted Apr 16, 2013

Inventors: James A. Hayward (Stony Brook, NY); Ming-Hwa Liang (Stony Brook, NY); Stephane Shu-Kin-So (Middle Island, NY)

Assignee: APDN (B.V.I.), Inc.

Abstract

Claims (122)

1. The method for authenticating a tablet comprising the steps of:

providing an optical reporter marker, the optical reporter marker having at least one light emitting upconverting phosphor particle linked by a linking group comprising an alkylene moiety, to a nucleic acid material, the nucleic acid material having a plurality of polymorphic double stranded (ds) DNA fragments, said polymorphic dsDNA fragments each having an artificial sequence obtained by digestion and ligation of extracted DNA,

applying the optical reporter marker to the tablet of interest,

detecting the optical reporter marker associated with the tablet with a light source,

obtaining a sample of the optical reporter marker from the tablet of interest for analysis,

analyzing the collected sample to detect the presence of the identifiable portion of the polymorphic (ds)DNA fragments linked to the upconverting phosphor particle by performing a polymerase chain reaction (PCR) on the collected sample comprising said non-heritable polymorphic (ds)DNA fragments using specific primers, thereby producing specific length amplicons; and

verifying that the tablet is genuine by detecting the specific length amplicons amplified from said polymorphic dsDNA fragments in said tablet.

2. The method of claim 1 , wherein the optical reporter marker has the composition of the formula I:

(cOpR)-[L-(NA)] m   I

wherein:

m is an integer greater than 1;

(cOpR) is a coated optical reporter particle, said COpR having an upconverting phosphor (UCP) material;

(NA) is a nucleic acid oligomer of detectable sequence, wherein (NA) is a double stranded DNA molecule having a length of between 40 base pairs and 1000 base pairs; and

L is the linking group having an alkylene moiety and being covalently bound to the coated optical reporter particle and to the nucleic acid oligomer.

3. The method of claim 2 , wherein (NA) is a double stranded DNA molecule having a length of between about 100 base pairs and about 800 base pairs.

4. The method of claim 2 , wherein L comprises an alkylene moiety, and having a first end covalently bound to the coated optical reporter particle and a second end covalently bound to the nucleic acid oligomer.

5. The method of claim 1 , wherein (UCP) is an upconverting phosphor particle of the formula:

Y x Yb y Er z O 2 S; or

Na(Y x Yb y Er z )F 4 ;

wherein:

x is from about 0.6 to about 0.95;

y is from about 0.05 to about 0.35; and

z is from about 0.1 to about 0.001.

6. The method of claim 2 , wherein L is of the formula:

-A-R 1 —B—

wherein:

R 1 is C 2-8 alkylene;

-A- is a group covalently bonded to the surface of the coated optical reporter; and

—B— is a group covalently bonded to the 3′ or 5′ end of the nucleic acid oligomer.

7. A method for authenticating a tablet comprising the steps of;

providing a tablet comprising an optical reporter marker, the optical reporter marker having at least one light emitting upconverting phosphor particle linked by a linking group comprising an alkylene moiety, to a nucleic acid taggant, the nucleic acid taggant having a plurality of polymorphic double stranded (ds) DNA fragments, said polymorphic dsDNA fragments each having an artificial sequence obtained by digestion and ligation of extracted DNA,

detecting the optical reporter marker associated with the tablet with a light source,

obtaining a sample of the optical reporter marker from the tablet for analysis,

analyzing the collected sample to detect the presence of the identifiable portion of the polymorphic (ds)DNA fragments linked to the upconverting phosphor particle by performing a polymerase chain reaction (PCR) on the collected sample comprising said polymorphic (ds)DNA fragments using specific primers, thereby producing specific length amplicons; and

verifying that the tablet is genuine by detecting the specific-length amplicons amplified from said polymorphic dsDNA fragments in said tablet.

8. The method of claim 7 , wherein the optical reporter marker has the composition of the formula I:

(cOpR)-[L-(NA)] m   I

wherein:

m is an integer greater than 1;

(cOpR) is a coated optical reporter particle, said COpR having an upconverting phosphor (UCP) material;

(NA) is a nucleic acid oligomer of detectable sequence, wherein (NA) is a double stranded DNA molecule having a length of between 40 base pairs and 1000 base pairs; and

L is the linking group comprising an alkylene moiety, and being covalently bound to the coated optical reporter particle and to the nucleic acid oligomer.

9. The method of claim 8 , wherein (NA) is a double stranded DNA molecule having a length of between about 100 base pairs and about 800 base pairs.

10. The method of claim 8 , wherein L comprises a C 2-8 alkylene moiety, and having a first end covalently bound to the coated optical reporter particle and a second end covalently bound to the nucleic acid oligomer.

11. The method of claim 8 , wherein (UCP) is an upconverting phosphor particle of the formula:

Y x Yb y Er z O 2 S; or

Na(Y x Yb y Er z )F 4 ;

wherein:

x is from about 0.6 to about 0.95;

y is from about 0.05 to about 0.35; and

z is from about 0.1 to about 0.001.

12. The method of claim 8 , wherein L is of the formula:

-A-R 1 —B—

wherein:

R 1 is C 2-8 alkylene;

-A- is a group covalently bonded to the surface of the coated optical reporter; and

—B— is a group covalently bonded to the 3′ or 5′ end of the nucleic acid oligomer.

13. The method of claim 12 , wherein -A- is —O—.

14. The method of claim 12 , wherein —R 1 — is —(CH 2 ) n — and wherein n is from 2 to 8.

15. The method of claim 12 , wherein —B— is:

—S—;

—O—;

—NR a —;

—S—(CH 2 ) p —;

—O—(CH 2 ) p —;

—NR a —(CH 2 ) p —;

—S—(CH 2 ) q —C(O)—NR a —(CH 2 ) p —;

—O—(CH 2 ) q —C(O)—NR a —(CH 2 ) p —;

—NR a —(CH 2 ) q —C(O)—NR a —(CH 2 ) p —;

—S—C(O)—(CH 2 ) r —C(O)—NR a —(CH 2 ) p —;

—O—C(O)—(CH 2 ) r —C(O)—NR a —(CH 2 ) p —; or

—NR a —C(O)—(CH 2 ) r —C(O)—NR a —(CH 2 ) p —;

wherein:

p is from 2 to 8;

q is from 1 to 8;

r is from 2 to 8; and

each R a is independently hydrogen or C 1-6 alkyl.

16. The method of claim 12 , wherein —B— is:

—S—(CH 2 ) q —C(O)—NR a —(CH 2 ) p or

—NR a —C(O)—(CH 2 ) r —C(O)—NR a —(CH 2 ) p —;

p is from 2 to 6;

q is from 1 to 3; and

r is 2 or 3.

17. The method of claim 12 , wherein —B— is:

—S—CH 2 —C(O)—NH—(CH 2 ) 6 —; or

—NH—C(O)—(CH 2 ) 3 —C(O)—NH—(CH 2 ) 6 —.

18. The method of claim 8 , wherein the cOpR is coated with silica.

19. The method of claim 18 , wherein cOpR is a coated optical reporter particle having an upconverting phosphor (UCP) material comprising Yttrium oxysulfide.

20. The method of claim 7 , wherein the optical reporter marker has the composition of the formula II:

(UCP)-[A-R 1 —X—R 2 —C(O)—NR a —R 3 -(DNA)] m   II

wherein:

m is an integer greater than 1;

UCP is an upconverting phosphor particle;

DNA is a single or double stranded deoxyribonucleic acid fragment;

-A- is a group capable of covalently bonding to the surface of the Upconverting phosphor particle;

R 1 is C 2-8 alkylene,

R 2 is C 1-8 alkylene or —C(O)—C 1-8 alkylene-;

—X— is —O—, —S— or —NR a —;

R 3 is C 2-8 alkylene; and

R a is hydrogen or C 1-6 alkyl.

21. The method of claim 20 , wherein the optical reporter marker has the composition of the formula IV:

(UCP)-[O—(CH 2 ) s —S—(CH 2 ) t —C(O)—NH—(CH 2 ) v -(DNA)] m   IV

wherein:

s is from 2 to 6;

v is from 2 to 6; and

t is from 1 to 3.

22. The method of claim 20 , wherein the optical reporter marker has the composition of the formula V:

(UCP)-[O—(CH 2 ) s —NH—C(O)—(CH 2 ) u —C(O)—NH—(CH 2 ) v -(DNA)] m   V

wherein:

s is from 2 to 6;

v is from 2 to 6; and

u is 2 or 3.

23. The method of claim 20 , wherein the optical reporter marker has the composition of the formula VI:

(UCP)-[O—(CH 2 ) 3 —S—CH 2 —C(O)—NH—(CH 2 ) 6 -(DNA)] m   VI.

24. The composition of claim 20 , wherein the composition is of the formula VII:

(UCP)-[O—(CH 2 ) 3 —NH—C(O)—(CH 2 ) 3 —C(O)—NH—(CH 2 ) 6 -(DNA)] m   VII.

25. The composition of claim 8 , wherein the cOpR comprises a visually detectable light emitting material selected from the group consisting of a fluorescent dye, a upconverting phosphor, a rare earth doped-ceramic powder, and a quantum dot.

26. The composition of claim 25 , where said light emitting materials are excitable by UV or infrared light.

27. The method of claim 8 , wherein the cOpR comprises at least one electromagnetic radiation emitting material.

28. The composition of claim 27 , where the electromagnetic radiation emitting material is detectable by a device which provides a source selected from the group consisting of an infrared radiation source, magnetic field source, a quantum dot or electromagnetic pulse source.

29. The method of claim 2 , wherein verifying that the tablet is genuine by detecting said polymorphic dsDNA fragments in said tablet, comprises analyzing the specific lengths of said specific length amplicons with capillary electrophoresis.

30. The method of claim 12 , wherein verifying that the tablet is genuine by detecting said polymorphic dsDNA fragments in said article, comprises analyzing the specific length amplicons amplified from said polymorphic (ds)DNA fragments with capillary electrophoresis.

Assignments (4)

Continuity (8)