Far-red dye probe formulations
Disclosed are formulations, including both liquid and lyophilized formulations, comprising a far-red dye probe and a non-linear surfactant or foamban. Also disclosed are related methods for preparing a lyophilized far-red dye probe formulation as well as related kits and diagnostic products.
1. A stabilized far-red dye probe formulation comprising:
a far-red dye probe comprising a far-red dye conjugated to a carrier molecule;
a non-linear surfactant at a concentration from about 0.5% (v/v) to about 20% (v/v); and
at least one buffering agent;
wherein the formulation is an aqueous solution, and further wherein:
(i) the non-linear surfactant is selected from the group consisting of a polyoxyethylene sorbitan fatty acid ester and digitonin; or
(ii) the far-red dye probe in the formulation has a relative fluorescence, in relative fluorescence units (RFU), and the relative fluorescence of the far-red dye probe is decreased by no more than 20% after 30 days of storage.
2. The formulation of claim 1 , wherein the far-red dye is a far-red cyanine dye.
3. The formulation of claim 2 , wherein the far-red cyanine dye is selected from the group consisting of cyanine 5 and cyanine 5.5.
4. The formulation of claim 1 , wherein the non-linear surfactant is selected from the group consisting of a polyoxyethylene sorbitan fatty acid ester and digitonin.
5. The formulation of claim 4 , wherein the non-linear surfactant is a polyoxyethylene sorbitan fatty acid ester selected from the group consisting of polysorbate 20, polysorbate 40, and polysorbate 60.
6. The formulation of claim 1 , wherein the far-red dye probe in the formulation has a relative fluorescence, in relative fluorescence units (RFU), and the relative fluorescence of far-red dye probe is decreased by no more than 20% after 30 days of storage.
7. The formulation of claim 1 , wherein the non-linear surfactant concentration is from about 0.5% (v/v) to about 10% (v/v), from about 1% (v/v) to about 20% (v/v), or from about 1% (v/v) to about 10% (v/v).
8. The formulation of claim 1 , wherein the at least one buffering agent is Tris.
9. The formulation of claim 8 , wherein the Tris buffering agent is present at a concentration of from about 5 mM to about 50 mM.
10. The formulation of claim 1 , wherein the carrier molecule is a nucleic acid.
11. The formulation of claim 10 , wherein the nucleic acid carrier molecule is an RNA.
12. The formulation of claim 1 , wherein the far-red dye probe further comprises a quencher.
13. The formulation of claim 12 , wherein the far-red dye probe is selected from the group consisting of a molecular torch, a molecular beacon, and a TaqMan probe.
14. The formulation of claim 10 , further comprising a first amplification oligomer,
wherein the far-red dye probe comprises a target-hybridizing sequence that specifically binds to a first sequence contained within a target region of a target nucleic acid,
wherein the first amplification oligomer comprises a target-hybridizing sequence that specifically binds to a second sequence contained within said target region, and
wherein the first amplification oligomer is configured to produce, in an amplification assay comprising the target nucleic acid as a template, an amplification product containing said target region.
15. The formulation of claim 14 , further comprising a second amplification oligomer,
wherein the second amplification oligomer comprises a target-hybridizing sequence that specifically binds to a third sequence contained within said target region, and
wherein the first and second amplification oligomers are configured to amplify said target region in multiple cycles of the amplification assay.
16. A stabilized far-red dye probe formulation comprising:
a far-red dye probe comprising a far-red dye conjugated to a carrier molecule,
wherein the carrier molecule is a nucleic acid;
a non-linear surfactant at a concentration from about 0.5% (v/v) to about 20% (v/v); and
at least one buffering agent; and
a first amplification oligomer;
wherein the formulation is an aqueous solution;
wherein the far-red dye probe comprises a target-hybridizing sequence that specifically binds to a first sequence contained within a target region of a target nucleic acid;
wherein the first amplification oligomer comprises a target-hybridizing sequence that specifically binds to a second sequence contained within said target region; and
wherein the first amplification oligomer is configured to produce, in an amplification assay comprising the target nucleic acid as a template, an amplification product containing said target region; and
wherein the first amplification oligomer is a promoter-based amplification oligomer further comprising a promoter sequence located 5′ to the first target-hybridizing sequence.
17. The formulation of claim 14 , further comprising one or more nucleotide triphosphates suitable for performing said amplification assay.
18. The formulation of claim 14 , further comprising one or more salts or co-factors suitable for performing said amplification assay.
19. A method of preparing a stabilized, lyophilized far-red dye probe formulation, the method comprising:
providing a stabilized far-red dye probe formulation comprising:
a far-red dye probe comprising a far-red dye conjugated to a carrier molecule;
a non-linear surfactant at a concentration from about 0.5% (v/v) to about 20% (v/v); and
at least one buffering agent;
wherein the formulation is an aqueous solution; and
lyophilizing the aqueous solution to form the lyophilized far-red dye probe formulation.
20. A stabilized, lyophilized far-red dye probe formulation prepared by the method of claim 19 .
21. The formulation of claim 1 , wherein the non-linear surfactant concentration is from about 1% (v/v) to about 10% (v/v) and the far-red dye probe in the formulation has a relative fluorescence, in relative fluorescence units (RFU), and the relative fluorescence of far-red dye probe is decreased by no more than 20% after 30 days of storage.
22. The formulation of claim 21 , wherein the non-linear surfactant concentration is from about 1% (v/v) to about 3% (v/v).
23. The formulation of claim 22 , wherein the non-linear surfactant concentration is about 1% (v/v), or about 1.24% (v/v), or about 1.5% (v/v), or about 1.6% (v/v), or about 3% (v/v).
24. The formulation of claim 6 , wherein the far-red dye is a far-red cyanine dye.
25. The formulation of claim 24 , wherein the non-linear surfactant concentration is from about 1% (v/v) to about 10% (v/v).
26. The formulation of claim 25 , wherein the non-linear surfactant concentration is from about 1% (v/v) to about 3% (v/v).
27. The method of claim 19 , wherein the non-linear surfactant is selected from the group consisting of a polyoxyethylene sorbitan fatty acid ester and digitonin.
28. The method of claim 19 , wherein the far-red dye probe in the stabilized far-red dye probe formulation has a relative fluorescence, in relative fluorescence units (RFU), and the relative fluorescence of the far-red dye probe is decreased by no more than 20% after 30 days of storage.
29. The formulation of claim 1 , wherein the non-linear surfactant is a polyoxyethylene sorbitan fatty acid ester.