Methods and articles for strand-specific polynucleotide detection with conjugated polymers
View Patent ↗The invention further relates to conjugated polymers, and methods, articles and compositions employing them as described herein. In some aspects, the invention relates to methods, articles and compositions for the detection and analysis of biomolecules in a sample. Provided assays include those determining the presence of a target biomolecule in a sample or its relative amount, or the assays may be quantitative or semi-quantitative. The methods can be performed on a substrate. The methods can be performed in an array format on a substrate, which can be a sensor. In some embodiments, detection assays are provided employing sensor biomolecules that do not comprise a fluorophore that can exchange energy with the cationic multichromophore. In some aspects biological assays are provided in which energy is transferred between one or more of the multichromophore, a label on the target biomolecule, a label on the sensor biomolecule, and/or a fluorescent dye specific for a polynucleotide, in all permutations. The multichromophore may interact at least in part electrostatically with the sensor and/or the target, and an increase in energy transfer with the polymer may occur upon binding of the sensor and the target. Other variations of the inventions are described further herein.
1. A detection method comprising:
providing a sample that is suspected of containing a target biomolecule;
providing a multichromophore that comprises a conjugated polymer, wherein said polymer comprises a plurality of low bandgap repeat units sufficient for the polymer to form an excited state when the polymer is contacted with incident light in the region of about 450 nm to about 1000 nm, and wherein said multichromophore has a detectable emission upon excitation and wherein the multichromophore can bind to the target biomolecule;
providing a sensor biomolecule capable of selectively binding to the target biomolecule, wherein the sensor is conjugated to a substrate;
contacting the substrate with a solution comprising the sample; and
detecting whether target biomolecule, if present, has bound to the sensor by applying a light source that can excite the multichromophore and detecting whether light is emitted in the vicinity of the substrate.
2. The method according to claim 1 , wherein the sensor biomolecule is selected from the group consisting of a peptide, a polynucleotide, a polysaccharide, and a protein.
3. The method according to claim 1 , wherein the sensor biomolecule is not conjugated to a fluorescently detectable label that can receive energy from an excited state of the multichromophore.
4. The method according to claim 1 , wherein the target biomolecule is selected from the group consisting of a peptide, a polynucleotide, a polysaccharide, and a protein.
5. The method according to claim 1 , wherein the sensor biomolecule comprises a protein.
6. The method according to claim 1 , wherein the target biomolecule comprises a protein.
7. The method according to claim 1 , wherein one of said target biomolecule and said sensor biomolecule comprises signaling chromophore that can receive energy from an excited state of said multichromophore.
8. The method according to claim 7 , wherein said target molecule comprises a signaling chromophore that can receive energy from an excited state of said multichromophore.
9. The method according to claim 7 , wherein said sensor biomolecule comprises a signaling chromophore that can receive energy from an excited state of said multichromophore.
10. The method according to claim 1 , wherein said substrate is a particle.
11. The method according to claim 1 , wherein the method further comprises quantitating the target biomolecule in the sample.
12. The method according to claim 11 , wherein the method further comprises washing the substrate prior to said detecting.
13. A detection method comprising:
providing a sample that is suspected of containing a target biomolecule;
providing a multichromophore that comprises a conjugated polymer, wherein said polymer comprises a plurality of low bandgap repeat units sufficient for the polymer to form an excited state when the polymer is contacted with incident light in the region of about 450 nm to about 1000 nm and wherein the multichromophore can bind to the target biomolecule;
wherein the target biomolecule is attached to a fluorescently detectable label that can receive energy from an excited state of the multichromophore;
providing a sensor biomolecule capable of recognition pairing with the target biomolecule, wherein the sensor biomolecule is conjugated to a substrate;
contacting the substrate with the solution comprising the sample under conditions in which the sensor biomolecule can pair with the target if present;
contacting the substrate with the multichromophore under conditions wherein the multichromophore can bind to the target biomolecule if present; and
detecting whether the target has bound to the sensor biomolecule by detecting whether energy has been transferred from the multichromophore to the fluorescently detectable label.
14. The method according to claim 13 , wherein the sensor biomolecule is selected from the group consisting of a peptide, a polynucleotide, a polysaccharide, and a protein.
15. The method according to claim 13 , wherein the sensor biomolecule is not conjugated to a fluorescently detectable label that can receive energy from an excited state of the multichromophore.
16. The method according to claim 13 , wherein the target biomolecule is selected from the group consisting of a peptide, a polynucleotide, a polysaccharide, and a protein.
17. The method according to claim 13 , wherein the sensor biomolecule comprises a protein.
18. The method according to claim 13 , wherein the target biomolecule comprises a protein.
19. The method according to claim 13 , wherein the substrate is not chemically conjugated to a fluorescently detectable label that can receive energy from an excited state of the chromophore.
20. The method according to claim 13 , wherein said substrate is a particle.
21. The method according to claim 13 , wherein the method further comprises quantitating the target biomolecule in the sample.
22. The method according to claim 13 , wherein the method further comprises washing the substrate prior to said detecting.