Optical reporter compositions
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.
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 x Yb y Er z ) F 4 ;
(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-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.
4. The composition of claim 3 , wherein -A- is —O—.
5. The composition of claim 3 , wherein —R 1 — is —(CH 2 ) n - and wherein n is from 2 to 8.
6. The composition of claim 3 , 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.
7. The composition of claim 3 , wherein —B— is:
—S—(CH 2 ) q —C(O)—NR a —(CH 2 ) p or
—N R a —C(O)—(CH 2 ) r —C(O)—NR a —(CH 2 ) 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—CH 2 —C(O)—NH—(CH 2 ) 6 —; or
—NH—C(O)—(CH 2 ) 3 —C(O)—NH—(CH 2 ) 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.