Flexible carbohydrate-bearing polymer

Abstract

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 - 23 . (canceled) 
     
     
         24 . A method of synthesizing a flexible water-soluble polymer comprising polymerized residues of first monomer units, the first monomer unit residues bearing pendant carbohydrate or carbohydrate mimetic moieties and pendant moieties which are one of a 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 one of a proximity based signal generating/modulating moiety pair, the method comprising one of the following procedures:
 (a) polymerising monomer units each bearing a pendant carbohydrate or carbohydrate mimetic moiety and a pendant proximity based signal generating/modulating moiety and optionally third monomer units;   (b) co-polymerising first monomer units each bearing a pendant carbohydrate or carbohydrate mimetic moiety and second monomer units each bearing a pendant proximity based signal generating/modulating moiety and optionally third monomer units;   (c) polymerising monomer units each bearing a pendant carbohydrate or carbohydrate mimetic moiety and a pendant functional group for linking to an proximity based signal generating/modulating moiety and optionally third monomer units, then reacting the monomer unit residues with the proximity based signal generating/modulating moieties;   (d) co-polymerising first monomer units each bearing a pendant carbohydrate or carbohydrate mimetic moiety and second monomer units each bearing a pendant functional group for linking to an proximity based signal generating/modulating moiety and optionally third monomer units, then reacting the second monomer unit residues with the proximity based signal generating/modulating moieties;   (e) polymerising monomer units each bearing a pendant functional group for linking to a carbohydrate or carbohydrate mimetic moiety and a pendant different functional group for linking to an proximity based signal generating/modulating moiety and optionally third monomer units, then reacting the monomer unit residues with the carbohydrate or carbohydrate mimetic moieties and proximity based signal generating/modulating moieties; or   (f) co-polymerising first monomer units each bearing a pendant functional group for linking to a carbohydrate or carbohydrate mimetic moiety and second monomer units each bearing a pendant different functional group for linking to an proximity based signal generating/modulating moiety and optionally third monomer units, then reacting the first monomer unit residues with the carbohydrate or carbohydrate mimetic moieties and the second monomer unit residues with proximity based signal generating/modulating moieties.   
     
     
         25 . A method as claimed in  claim 24 , wherein the monomer units are reacted by addition polymerization, condensation polymerization, ring opening polymerization or atom transfer radical polymerization. 
     
     
         26 . A method as claimed in  claim 24 , wherein the monomer units and/or the first monomer units comprise allyl or vinyl containing derivatives of carbohydrate or carbohydrate mimetic moieties. 
     
     
         27 . A method as claimed in  claim 24 , wherein the monomer units and/or the second monomer units comprise double bond-containing molecules containing an amine functional group. 
     
     
         28 . A method as claimed in  claim 24 , wherein the first monomer units are present in the polymerization reaction mixture in an amount of 20 to 70 mol % or 70 to 90 mol % and/or the second monomer units are present in the reaction mixture in an amount of 5 to 15 mol %. 
     
     
         29 . A method as claimed in  claim 24 , wherein the carbohydrate moieties are selected from optionally derivatised mannose, maltose, isomaltose, glucose, sophorose and/or 2-acetylglucosamine. 
     
     
         30 . A method as claimed in  claim 24 , wherein the carbohydrate mimetic moieties are carbohydrate mimetic peptide moieties. 
     
     
         31 . A method as claimed in  claim 24 , 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 and/or the second monomer units. 
     
     
         32 . A method as claimed in  claim 24 , wherein the proximity based signal generating/modulating moieties are energy donors or energy acceptors. 
     
     
         33 . A method as claimed in  claim 32 , wherein the energy acceptors are HMCV-1. 
     
     
         34 . A method as claimed in  claim 24 , wherein the third monomer units do not bear carbohydrate or carbohydrate mimetic moieties or proximity based signal generating/modulating moieties. 
     
     
         35 . A method as claimed in  claim 24 , wherein the third monomer units are present in the reaction mixture in an amount of 0 to 80 mol %. 
     
     
         36 . A sensor as claimed in  claim 24 , wherein the third monomer units contain hydrophilic groups. 
     
     
         37 . A method as claimed in  claim 36 , wherein the third monomer units comprise 2-hydroxyethylacrylate, vinyl pyrrolidone, MMA, HEMA, vinyl alcohol and/or ethylene glycol. 
     
     
         38 . A method as claimed in  claim 24 , wherein the average polymer molecular weight is in the range of 20 to 250 kDa. 
     
     
         39 . A method as claimed in  claim 24 , wherein the polymer has a non-carbohydrate backbone. 
     
     
         40 . A method as claimed in  claim 24 , wherein the polymer is unbranched. 
     
     
         41 . A method as claimed in  claim 24 , wherein the first monomer unit residues and the second monomer unit residues are different in structure not just because of their pendant moieties. 
     
     
         42 . A method as claimed in  claim 24 , wherein the polymer has less than 10% double bonds in the backbone. 
     
     
         43 . A method as claimed in  claim 24 , wherein the first monomer units are allyl α-D-mannopyranoside, the second monomer units are labeled N-(3-aminopropyl)methacrylamide and the third monomer units are 2-hydroxyethylacrylate.