Optical reporter compositions

• US 8,372,648 B2
• Filed: 12/11/2007
• Issued: 02/12/2013
• Est. Priority Date: 04/16/2003
• Status: Active Grant

• Current Assignee: APDN (B.V.I.) Inc. (Applied DNA Sciences Incorporated)
• Original Assignee: APDN (B.V.I.) Inc. (Applied DNA Sciences Incorporated)
• Inventors: Kwok, Thomas, Liang, Ming-Hwa, Shu Kin So, Stephane
• Primary Examiner(s): Menon, Krishnan S
• Assistant Examiner(s): Fritchman, Rebecca M
• Application Number:
  US11/954,030
• Publication Number:
  US 20080299667A1
• Time in Patent Office: 1,890 Days
• Field of Search: 436/91, 436/94, 435/5, 435/6.11, 435/6.18, 536/25.3
• US Class Current: 436/94
• CPC Class Codes: B82Y 15/00 Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
• CPC Class Codes: C08K 5/09 Carboxylic acids; Metal salts thereof; Anhydrides thereof
• CPC Class Codes: C09D 11/03 characterised by features other than the chemical nature of the binder
• CPC Class Codes: C09D 7/63 organic
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 2563/185 Nucleic acid dedicated to use as a hidden marker/bar code, e.g. inclusion of nucleic acids to mark art objects or animals
• CPC Class Codes: Y10S 977/774 Exhibiting three-dimensional carrier confinement, e.g. quantum dots
• CPC Class Codes: Y10S 977/92 Detection of biochemical
• CPC Class Codes: Y10T 436/14 Heterocyclic carbon compound [i.e. , O, S, N, Se, Te, as only ring hetero atom]
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

This invention provides compositions that have a light emitting reporter linked to biomolecules, preferably, nucleotide oligomers. The light reporter particles are silylated and functionalized to produce a coated light reporter particle, prior to covalently linking the biomolecules to the light reporter particle. The light reporter particles of the invention can be excited by a light excitation source such as UV or IR light, and when the biomolecule is DNA, the attached DNA molecule(s) are detectable by amplification techniques such as PCR.

Claims

1. A 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 comprising an upconverting phosphor (UCP) of the formula Na (Y_xYb_yEr_z) F₄;

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

L is a linking group covalently bound to the coated optical reporter particle and to the nucleic acid oligomer, said linking group L comprises an alkylene moiety having a first end covalently bound to the coated optical reporter particle and a second end covalently bound to the nucleic acid oligomer.

2. The composition of claim 1, 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.

3. The composition of claim 1, wherein L is of the formula:

-A-R¹—

B—

wherein;

R¹ is C_2-8alkylene;

-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.

4. The composition of claim 3, wherein -A- is —

O—

.

5. The composition of claim 3, wherein —

R¹—

is —

(CH₂)_n- and wherein n is from 2 to 8.

6. The composition of claim 3, wherein —

B—

is;

—

S—

;

—

O—

;

—

NR^a—

;

—

S—

(CH₂)_p—

;

—

O—

(CH₂)_p—

;

—

NR^a—

(CH₂)_p—

;

—

S—

(CH₂)_q—

C(O)—

NR^a—

(CH₂)_p—

;

—

O—

(CH₂)_q—

C(O)—

NR^a—

(CH₂)_p—

;

—

NR^a—

(CH₂)_q—

C(O)—

NR^a—

(CH₂)_p—

;

—

S—

C(O)—

(CH₂)_r—

C(O)—

NR^a—

(CH₂)_p—

;

—

O—

C(O)—

(CH₂)_r—

C(O)—

NR^a—

(CH₂)_p—

;

or

—

NR^a—

C(O)—

(CH₂)_r—

C(O)—

NR^a—

(CH₂)_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_1-6alkyl.

7. The composition of claim 3, wherein —

B—

is;

—

S—

(CH₂)_q—

C(O)—

NR^a—

(CH₂)_p or

—

N R^a—

C(O)—

(CH₂)_r—

C(O)—

NR^a—

(CH₂)_p—

;

wherein p, q, r and R^a are as recited in claim 6.

8. The composition of claim 7, wherein:

p is from 2 to 6;

q is from 1 to 3; and

r is 2 or 3.

9. The composition of claim 3, wherein —

B—

is;

—

S—

CH₂—

C(O)—

NH—

(CH₂)₆—

;

or

—

NH—

C(O)—

(CH₂)₃—

C(O)—

NH—

(CH₂)₆—

.

10. The composition of claim 1, wherein the cOpR is coated with silica.

11. The composition of claim 10, wherein the silica comprises at least one Si—

O bond.

12. The composition of claim 1, wherein the NA is double stranded DNA molecule identifiable by real time PCR without DNA sequencing.

13. The composition of claim 1, wherein the NA is a DNA molecule having a length of between 80 base pairs and 500 base pairs.

14. The composition of claim 1, wherein said nucleic acid oligomer is identifiable by a method comprising polymerase chain reaction amplification while said oligomer is covalently attached to said coated optical reporter particle.

15. The composition of claim 14, wherein said polymerase chain reaction amplification produces one or more amplicons and the size of said one or more amplicons is determined by a method comprising capillary electrophoresis.