Flexible carbohydrate-bearing polymer
View Patent ↗A sensor for the detection or measurement of a carbohydrate analyte in fluid comprises components of a competitive binding assay the readout of which is a detectable or measurable optical signal retained by a material that permits diffusion of the analyte but not the assay components, the assay components comprising: a carbohydrate binding molecule labelled with one of a proximity based signal generating/modulating moiety pair; and a carbohydrate analogue capable of competing with the analyte for binding to the carbohydrate binding molecule, the carbohydrate analogue being a flexible water-soluble polymer comprising: polymerized or co-polymerised residues of monomer units, the monomer unit residues bearing pendant carbohydrate or carbohydrate mimetic moieties and pendant moieties which are the other of the proximity based signal generating/modulating moiety pair.
1. A sensor for the detection or measurement of a carbohydrate analyte in fluid, the sensor comprising components of a competitive binding assay the readout of which is a detectable or measurable optical signal retained by a material that permits diffusion of the analyte but not the assay components, the assay components comprising:
a carbohydrate binding molecule labeled with one of a proximity based signal generating/modulating moiety pair wherein the carbohydrate binding molecule specifically binds to the carbohydrate analyte; and
a carbohydrate analogue capable of competing with the analyte for binding to the carbohydrate binding molecule, the carbohydrate analogue being a flexible water-soluble polymer comprising:
polymerized residues of monomer units, the monomer unit residues bearing pendant carbohydrate or carbohydrate mimetic moieties and pendant moieties which are the other of the proximity based signal generating/modulating moiety pair; and/or
co-polymerised residues of first monomer units and second monomer units, the first monomer unit residues bearing pendant carbohydrate or carbohydrate mimetic moieties and the second monomer unit residues bearing pendant moieties which are the other of the proximity based signal generating/modulating moiety pair;
where said polymer has a non-carbohydrate backbone.
2. A sensor as claimed in claim 1 , comprising a copolymer of said first and second monomer units polymerized from a reaction mixture, wherein the second monomer units are present in the reaction mixture in an amount of 5 to 15 mol %.
3. A sensor as claimed in claim 1 , wherein the carbohydrate moieties are selected from optionally derivatised mannose, maltose, isomaltose, glucose, sophorose and/or 2-acetylglucosamine.
4. A sensor as claimed in claim 1 , wherein the proximity based signal generating/modulating moieties are linked to amine, acid, alcohol, alkyne, azide and/or sulphone functional groups of the monomer units or the second monomer units.
5. A sensor as claimed in claim 1 , wherein the proximity based signal generating/modulating moieties are energy donors or energy acceptors.
6. A sensor as claimed in claim 1 , further comprising polymerized residues of third monomer units not bearing carbohydrate or carbohydrate mimetic moieties or proximity based signal generating/modulating moieties.
7. A sensor as claimed in claim 6 , wherein the third monomer units are present in the reaction mixture in an amount of 0 to 80 mol %.
8. A sensor as claimed in claim 6 , wherein the third monomer units contain hydrophilic groups.
9. A sensor as claimed in claim 8 , wherein the third monomer units comprise 2-hydroxyethylacrylate, vinyl pyrrolidone, MMA, HEMA, vinyl alcohol and/or ethylene glycol.
10. A sensor as claimed in claim 1 , wherein the average polymer molecular weight is in the range of 20 to 250 kDa.
11. A sensor as claimed in claim 1 , wherein the carbohydrate analyte is glucose and the polymer is capable of competing with glucose at physiological calcium concentrations.
12. A sensor as claimed in claim 11 , wherein the assay is capable of measuring blood glucose at concentrations over at least part of the range of 0 to 35 mM glucose.
13. A sensor as claimed in claim 1 , wherein the carbohydrate binding molecule is a lectin.
14. A sensor as claimed in claim 13 , wherein the carbohydrate binding molecule is mannose binding lectin or Concanavalin A.
15. A sensor as claimed in claim 1 , wherein the components of the assay are retained by a shell or matrix material.
16. A sensor as claimed in claim 15 , wherein the retaining material is biodegradable.
17. A method of preparing a sensor as claimed in claim 1 , the method comprising at least one of phase separation (coacervation), solvent evaporation, extraction, spray drying, spray coating, spray chilling, rotary disk atomisation, fluid bed coating, coextrusion and pan coating.
18. A method of detecting a carbohydrate analyte using a sensor as claimed in claim 1 , comprising implantation of the sensor into the skin of a mammal and transdermal detection or measurement of carbohydrate analyte using external optical means.
19. A method of detecting a carbohydrate analyte using a sensor as claimed in claim 1 , comprising transdermal detection or measurement of a carbohydrate analyte using external optical means by illumination of a said sensor present in or below the skin of a mammal.
20. A sensor as claimed in claim 1 , wherein the polymer is unbranched.
21. A sensor as claimed in claim 1 , wherein the first monomer unit residues and the second monomer unit residues are different in structure not just because of their pendant moieties.
22. A sensor as claimed in claim 1 , wherein the polymer has less than 10% double bonds in the backbone.
23. A sensor as claimed in claim 1 , wherein the polymer is formed by addition polymerization.
24. A sensor as claimed in claim 1 , wherein the polymer comprises polymerized residues of allyl α-D-mannopyranoside, 2-hydroxyethylacrylate and labeled N-(3-aminopropyl)methacrylamide.