Sonstiges: |
- Nachgewiesen in: USPTO Patent Grants
- Sprachen: English
- Patent Number: 11034,840
- Publication Date: June 15, 2021
- Appl. No: 16/667250
- Application Filed: October 29, 2019
- Assignees: BECTON, DICKINSON AND COMPANY (Franklin Lakes, NJ, US)
- Claim: 1. A method of evaluating a sample for the presence of a target analyte, the method comprising: (a) contacting the sample with an aggregation-resistant polymeric dye conjugate that specifically binds the target analyte to produce a labelling composition contacted sample, wherein the polymeric dye conjugate comprises: (i) a water soluble light harvesting multichromophore comprising a conjugated segment having the structure of formula (I): [chemical expression included] wherein: F 1 is a fused tricyclic co-monomer substituted with a water soluble group (WSG); M 1 is an aryl or heteroaryl co-monomer; n is an integer from 1 to 100,000; and * denotes a site for covalent attachment to the unsaturated backbone of a conjugated polymer or an end group; wherein at least one of F 1 and M 1 is substituted with a branched non-ionic water soluble group (WSG) comprising two or more water soluble polymers each having 6-50 monomeric units; and (ii) a specific binding member; and (b) assaying the labelling composition contacted sample for the presence of a polymeric dye conjugate-target analyte binding complex to evaluate whether the target analyte is present in the sample, wherein the multichromophore has one of the following formulae: [chemical expression included] wherein: each B 1 and B 2 are independently a branching group selected from the group consisting of CH, N, C(═O)N, SO 2 N, a tri-substituted aryl group, a tetra-substituted aryl group, and a tri-substituted heteroaryl group; each W 1 is independently a water soluble polymer comprising 6-24 monomeric units; T 3 is an optional linker to the fused 6-5-6 tricyclic co-monomer; and each p and q are independently 0 or 1, wherein if present, each T 1 and each T 2 are independently a linker, wherein each T 1 is independently selected from the group consisting of (CH 2) n —O—, —O—(CH 2) n —, —(CH 2) n —, and —O—.
- Claim: 2. The method according to claim 1 , wherein the polymeric dye conjugate further comprises a signaling chromophore covalently linked to the multichromophore in energy-receiving proximity therewith.
- Claim: 3. The method according to claim 1 , further comprising contacting the sample with a second specific binding member that is support bound and specifically binds the target analyte.
- Claim: 4. The method according to claim 3 , wherein the support comprises a magnetic particle.
- Claim: 5. The method according to claim 1 , wherein the target analyte is associated with a cell.
- Claim: 6. The method according to claim 5 , wherein the target analyte is a cell surface marker of the cell.
- Claim: 7. The method according to claim 6 , wherein the cell surface marker is selected from the group consisting of a cell receptor and a cell surface antigen.
- Claim: 8. The method according to claim 7 , wherein the target analyte is an intracellular target, and the method further comprises lysing the cell.
- Claim: 9. The method according to claim 1 , wherein the method further comprises flow cytometrically analyzing the fluorescently labelled target analyte.
- Claim: 10. A method of labelling a target molecule, the method comprising: contacting the target molecule with a polymeric dye to produce a labelled target molecule, wherein the polymeric dye comprises: a water soluble light harvesting multichromophore comprising a conjugated segment having the structure of formula (I): [chemical expression included] wherein: F 1 is a fused tricyclic co-monomer substituted with a water soluble group (WSG); M 1 is an aryl or heteroaryl co-monomer; n is an integer from 1 to 100,000; and * denotes a site for covalent attachment to the unsaturated backbone of a conjugated polymer or an end group; wherein at least one of F 1 and M 1 is substituted with a branched non-ionic water soluble group (WSG) comprising two or more water soluble polymers each having 6-50 monomeric units; and a conjugation tag that covalently links to the target molecule, wherein the multichromophore has one of the following formulae: [chemical expression included] wherein: each B 1 and B 2 are independently a branching group selected from the group consisting of CH, N, C(═O)N, SO 2 N, a tri-substituted aryl group, a tetra-substituted aryl group, and a tri-substituted heteroaryl group; each W 1 is independently a water soluble polymer comprising 6-24 monomeric units; T 3 is an optional linker to the fused 6-5-6 tricyclic co-monomer; and each p and q are independently 0 or 1, wherein if present, each T 1 and each T 2 are independently a linker, wherein each T 1 is independently selected from the group consisting of (CH 2) n —O—, —O—(CH 2) n —, —(CH 2) n —, and —O—.
- Claim: 11. The method according to claim 10 , wherein the polymeric dye further comprises a signaling chromophore covalently linked to the multichromophore in energy-receiving proximity therewith.
- Claim: 12. The method according to claim 10 , further comprising fluorescently detecting the labelled target molecule.
- Claim: 13. The method according to claim 10 , wherein the conjugation tag comprises a terminal functional group selected from an amino, a thiol, a hydroxyl, a hydrazine, a hydrazide, a azide, an alkyne, maleimide, iodoacetyl, amine, an active ester and a protein reactive group.
- Claim: 14. The method according to claim 10 , wherein the target molecule is a specific binding member.
- Claim: 15. The method according to claim 14 , wherein the specific binding member is an antibody.
- Claim: 16. The method according to claim 14 , wherein the specific binding member is an antibody fragment or binding derivative thereof.
- Claim: 17. A kit comprising: a water soluble light harvesting multichromophore comprising a conjugated segment having the structure of formula (I): [chemical expression included] wherein: F 1 is a fused tricyclic co-monomer substituted with a water soluble group (WSG); M 1 is an aryl or heteroaryl co-monomer; n is an integer from 1 to 100,000; and * denotes a site for covalent attachment to the unsaturated backbone of a conjugated polymer or an end group; wherein at least one of F 1 and M 1 is substituted with a branched non-ionic water soluble group (WSG) comprising two or more water soluble polymers each having 6-50 monomeric units; and a container, wherein the multichromophore has one of the following formulae: [chemical expression included] wherein: each B 1 and B 2 are independently a branching group selected from the group consisting of CH, N, C(═O)N, SO 2 N, a tri-substituted aryl group, a tetra-substituted aryl group, and a tri-substituted heteroaryl group; each W 1 is independently a water soluble polymer comprising 6-24 monomeric units; T 3 is an optional linker to the fused 6-5-6 tricyclic co-monomer; and each p and q are independently 0 or 1, wherein if present, each T 1 and each T 2 are independently a linker, wherein each T 1 is independently selected from the group consisting of (CH 2) n —O—, —O—(CH 2) n —, —(CH 2) n —, and —O—.
- Claim: 18. The kit according to claim 17 , further comprising one or more components selected from the group consisting of a polymeric tandem dye, a fluorophore, a specific binding member, a specific binding member conjugate, a cell, a support, a biocompatible aqueous elution buffer and instructions for use.
- Claim: 19. The kit according to claim 17 , wherein the multichromophore is covalently linked to a specific binding member.
- Claim: 20. The kit according to claim 17 , wherein multichromophore further comprises an acceptor signaling chromophore covalently linked to the multichromophore in energy-receiving proximity therewith.
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- Primary Examiner: Fang, Shane
- Attorney, Agent or Firm: Field, Bret E. ; Bozicevic, Field & Francis LLP
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