Polymer Probes And Methods

Abstract

A polymer detection probe is provided that includes a binding module that specifically binds to at least one polymer and a reporter module that is spectroscopically detectable. The binding module can be a carbohydrate-binding module (CBM). The reporter module can be a fluorescent protein. A complex is provided that includes a probe specifically bound to a pulp or paper product including at least one surface available lignocellulosic polymer. A pulp or paper product is provided that includes at least one surface available lignocellulosic polymer and at least one probe bound thereto. Methods are provided that employ a lignocellulosic probe. A method of detecting a lignocellulosic polymer or other type of polymer is provided. A method of determining the effectiveness of an industrial treatment on pulp or a paper product is also provided. A method of determining a physical property of pulp or a paper product is further provided.

1 - 28 . (canceled) 
     
     
         29 . A method of detecting a lignocellulosic polymer, the method comprising: contacting a lignocellulosic polymer detection probe with a biomass material; and
 measuring a property associated with the reporter module to determine the presence or absence of at least one lignocellulosic polymer in the biomass material based on specific binding of the probe to the at least one lignocellulosic polymer.   
     
     
         30 . The method of  claim 29 , wherein the property measured is fluorescence. 
     
     
         31 . The method of  claim 29 , further comprising calculating the amount of the at least one lignocellulosic polymer, determining the type of the at least one lignocellulosic polymer, or both. 
     
     
         32 . (canceled) 
     
     
         33 . (canceled) 
     
     
         34 . (canceled) 
     
     
         35 . (canceled) 
     
     
         36 . (canceled) 
     
     
         37 . (canceled) 
     
     
         38 . The method of  claim 31 , further comprising performing at least one treatment of the biomass material based on the amount of the at least one lignocellulosic polymer measured, the type of lignocellulosic polymer measured, or both. 
     
     
         39 . The method of  claim 31 , wherein the amount of lignocellulosic polymer measured correlates negatively or positively with at least one physical property of the biomass product. 
     
     
         40 . The method of  claim 39 , wherein the at least one physical property comprises burst index, drainage rate, tear index, tensile index, or internal bond strength, or any combination thereof. 
     
     
         41 . The method of  claim 31 , further comprising dosing at least one enzyme based on the amount of lignocellulosic polymer measured, the type of lignocellulosic polymer measured, or both. 
     
     
         42 . The method of  claim 31 , further comprising adjusting mill speed based on the amount of lignocellulosic polymer measured, the type of lignocellulosic polymer measured, or both. 
     
     
         43 . The method of  claim 31 , further comprising adjusting total water content of the biomass material based on the amount of lignocellulosic polymer measured, the type of lignocellulosic polymer measured, or both. 
     
     
         44 . The method of  claim 29 , wherein the probe comprises a plurality of probes. 
     
     
         45 . A method of determining the effectiveness of an industrial treatment on pulp or a paper product comprising:
 contacting a lignocellulosic polymer detection probe with a pulp or a paper product, said lignocellulosic polymer detection probe comprising a) a binding module that specifically binds to at least one lignocellulosic polymer and b) a reporter module that is spectroscopically detectable and wherein the binding module is a binding module polypeptide and the reporter module is a reporter module polypeptide that comprises a fluorescent protein and wherein the reporter module is not an antibody or a fragment thereof and wherein the binding module polypeptide is not an antibody or a fragment thereof;   detecting the specific binding of the probe to the pulp or the paper product;   
       calculating the amount of at least one lignocellulosic polymer on a surface of the pulp or the paper product; and
 determining the effectiveness of an industrial treatment on the pulp or paper product based on the amount of the at least one lignocellulosic polymer detected, wherein the fluorescent protein comprises Sirius, TagBFP, mTagBFP2, Azurite, EBFP2, mKalama1, Sirius, Sapphire, T-Sapphire, ECFP, mAmetrine, Cerulean, mCerulean3, SCFP3A, CyPet, mTurguoise, mTurquoise2, monomeric Midoriishi-Cyan, Aquamarine, eCFP, TagCFP, mTFP1, AmCyanl, EGFP, Emerald, Superfolder GFP, monomeric Azami Green, TurboGFP, TagGFP2, mUKG, mWasabi, Clover, mNeonGreen, eGFP, AcGFP1, ZGreen1, ZsYellow1, mBanana, EYFP, Topaz, Citrine, Venus, SYFP2, Ypet, IanRFP-deltaS83, mPapaya1, TagYFP, mOrange, mOrange2, monomeric Kusabira-Orange, mKOk, mKO2, mTangerine, mNectarine, TagRFP, TagRFP-T, mApple, mRuby, mRuby2, DsRed-Express2, DsRed-Express, tdTomato, DsRed-Monomer, DsRed-Monomer, DsRed2, AsRed2, mStrawberry, mCherry, HcRed1, FusionRed, mRaspberry, E2-Crimson, mPlum, HcRed-Tandem, mKate2, mNeptune, NirFP, TagRFP657, TagRFP675, iFP1.4, iRFP(iRFP713), iRFP670, iRFP682, iRFP702, iRFP720, iFP2.0, mIFP, mKeima Red, LSS-mKate1, LSS-mKate2, LSSmOrange, mBeRFP, PA-GFP, PATag RFP, Dendra2, Timer, PAmCherry, Kaede (green), Kaeda (red), KikGR1 (green), KikGR1 (red), PS-CFP2, mEos2 (green), mEos2 (red), mEos3.2 (green), mEos3.2(red), PSmOrange, Dropna, or any combination thereof and wherein the binding module is a carbohydrate-binding module (CBM) comprising CBM1, CBM2, CBM3, CBM3a CBM4, CBM5, CBM6, CBM9, CBM10, CBM11, CBM12, CBM14, CBM15, CBM17, CBM18, CBM19, CBM20, CBM21, CBM25, CBM27, CBM28, CBM33, CBM48, CBM49, or any combination thereof. 
 
     
     
         46 . (canceled) 
     
     
         47 . (canceled) 
     
     
         48 . The method of  claim 45 , wherein the lignocellulosic polymer detection probe is detectable at a distinct wavelength. 
     
     
         49 . A method of determining a physical property of pulp or a paper product comprising:
 contacting a lignocellulosic polymer detection probe with a pulp or a paper product, said lignocellulosic polymer detection probe comprising a) a binding module that specifically binds to at least one lignocellulosic polymer and b) a reporter module that is spectroscopically detectable and wherein the binding module is a binding module polypeptide and the reporter module is a reporter module polypeptide that comprises a fluorescent protein and wherein the reporter module is not an antibody or a fragment thereof and wherein the binding module polypeptide is not an antibody or a fragment thereof;   detecting the specific binding of the probe to the pulp or the paper product;   calculating the amount of at least one lignocellulosic polymer on a surface of the pulp or the paper product; and   determining at least one physical property of the pulp or paper product based on the amount of the at least one lignocellulosic polymer detected, wherein the fluorescent protein comprises Sirius, TagBFP, mTagBFP2, Azurite, EBFP2, mKalama1, Sirius, Sapphire, T-Sapphire, ECFP, mAmetrine, Cerulean, mCerulean3, SCFP3A, CyPet, mTurguoise, mTurquoise2, monomeric Midoriishi-Cyan, Aquamarine, eCFP, TagCFP, mTFP1, AmCyanl, EGFP, Emerald, Superfolder GFP, monomeric Azami Green, TurboGFP, TagGFP2, mUKG, mWasabi, Clover, mNeonGreen, eGFP, AcGFP1, ZGreen1, ZsYellow1, mBanana, EYFP, Topaz, Citrine, Venus, SYFP2, Ypet, IanRFP-deltaS83, mPapaya1, TagYFP, mOrange, mOrange2, monomeric Kusabira-Orange, mKOk, mKO2, mTangerine, mNectarine, TagRFP, TagRFP-T, mApple, mRuby, mRuby2, DsRed-Express2, DsRed-Express, tdTomato, DsRed-Monomer, DsRed-Monomer, DsRed2, AsRed2, mStrawberry, mCherry, HcRed1, FusionRed, mRaspberry, E2-Crimson, mPlum, HcRed-Tandem, mKate2, mNeptune, NirFP, TagRFP657, TagRFP675, iFP1.4, iRFP(iRFP713), iRFP670, iRFP682, iRFP702, iRFP720, iFP2.0, mIFP, mKeima Red, LSS-mKate1, LSS-mKate2, LSSmOrange, mBeRFP, PA-GFP, PATag RFP, Dendra2, Timer, PAmCherry, Kaede (green), Kaeda (red), KikGR1 (green), KikGR1 (red), PS-CFP2, mEos2 (green), mEos2 (red), mEos3.2 (green), mEos3.2(red), PSmOrange, Dropna, or any combination thereof and wherein the binding module is a carbohydrate-binding module (CBM) comprising CBM1, CBM2, CBM3, CBM3a CBM4, CBM5, CBM6, CBM9, CBM10, CBM11, CBM12, CBM14, CBM15, CBM17, CBM18, CBM19, CBM20, CBM21, CBM25, CBM27, CBM28, CBM33, CBM48, CBM49, or any combination thereof.   
     
     
         50 . The method of  claim 49 , wherein the at least one physical property comprises burst index, drainage rate, tear index, tensile index, or internal bond strength, or any combination thereof. 
     
     
         51 - 64 . (canceled) 
     
     
         65 . A method of detecting a polymer, the method comprising:
 contacting a polymer detection probe with a material, said polymer detection probe comprising a) a binding module that specifically binds to at least one polymer and b) a reporter module that is spectroscopically detectable and wherein the binding module is a binding module polypeptide and the reporter module is a reporter module polypeptide that comprises a fluorescent protein and wherein the reporter module is not an antibody or a fragment thereof and wherein the binding module polypeptide is not an antibody or a fragment thereof; and   measuring a property associated with the reporter module to determine the presence or absence of at least one polymer in the material based on specific binding of the probe to the at least one polymer, wherein the fluorescent protein comprises Sirius, TagBFP, mTagBFP2, Azurite, EBFP2, mKalama1, Sirius, Sapphire, T-Sapphire, ECFP, mAmetrine, Cerulean, mCerulean3, SCFP3A, CyPet, mTurguoise, mTurquoise2, monomeric Midoriishi-Cyan, Aquamarine, eCFP, TagCFP, mTFP1, AmCyanl, EGFP, Emerald, Superfolder GFP, monomeric Azami Green, TurboGFP, TagGFP2, mUKG, mWasabi, Clover, mNeonGreen, eGFP, AcGFP1, ZGreen1, ZsYellow1, mBanana, EYFP, Topaz, Citrine, Venus, SYFP2, Ypet, IanRFP-deltaS83, mPapaya1, TagYFP, mOrange, mOrange2, monomeric Kusabira-Orange, mKOk, mKO2, mTangerine, mNectarine, TagRFP, TagRFP-T, mApple, mRuby, mRuby2, DsRed-Express2, DsRed-Express, tdTomato, DsRed-Monomer, DsRed-Monomer, DsRed2, AsRed2, mStrawberry, mCherry, HcRed1, FusionRed, mRaspberry, E2-Crimson, mPlum, HcRed-Tandem, mKate2, mNeptune, NirFP, TagRFP657, TagRFP675, iFP1.4, iRFP(iRFP713), iRFP670, iRFP682, iRFP702, iRFP720, iFP2.0, mIFP, mKeima Red, LSS-mKate1, LSS-mKate2, LSSmOrange, mBeRFP, PA-GFP, PATag RFP, Dendra2, Timer, PAmCherry, Kaede (green), Kaeda (red), KikGR1 (green), KikGR1 (red), PS-CFP2, mEos2 (green), mEos2 (red), mEos3.2 (green), mEos3.2(red), PSmOrange, Dropna, or any combination thereof and wherein the binding module is a carbohydrate-binding module (CBM) comprising CBM1, CBM2, CBM3, CBM3a CBM4, CBM5, CBM6, CBM9, CBM10, CBM11, CBM12, CBM14, CBM15, CBM17, CBM18, CBM19, CBM20, CBM21, CBM25, CBM27, CBM28, CBM33, CBM48, CBM49, or any combination thereof.   
     
     
         66 . The method of  claim 65 , wherein the property measured is fluorescence. 
     
     
         67 . The method of  claim 65 , further comprising calculating the amount of the at least one polymer, determining the type of the at least one polymer, or both. 
     
     
         68 . The method of  claim 65 , wherein the material comprises a blood sample. 
     
     
         69 . The method of  68 , further comprising determining at least one of a blood antigen, type, group, and subgroup of the blood sample. 
     
     
         70 . The method of  claim 65 , wherein the probe comprises a plurality of probes. 
     
     
         71 . (canceled) 
     
     
         72 . (canceled) 
     
     
         73 . The method of  claim 29 , wherein the binding module polypeptide is fused directly to the reporter module polypeptide. 
     
     
         74 . The method of  claim 29 , wherein the binding module polypeptide is linked to the reporter polypeptide via a linker polypeptide. 
     
     
         75 . The method of  claim 29 , wherein the probe comprises an amino acid sequence of any one of SEQ ID NOS: 2, 6, 8, 10, and 12. 
     
     
         76 . The method of  claim 29 , wherein the binding module specifically binds to cellulose, hemicellulose, lignin, xylan, mannan, or any combination thereof. 
     
     
         77 . The method of  claim 29 , wherein the lignocellulosic polymer detection probe comprises a plurality of lignocellulosic polymer detection probes, each lignocellulosic polymer detection probe specifically binding to a different lignocellulosic polymer. 
     
     
         78 . The method of  claim 65 , wherein the binding module specifically binds to cellulose, hemicellulose, lignin, xylan, mannan, glucuronoxylan, arabinoxylan, glucomannan, xyloglucan, or any combination thereof or a linear fragment thereof, or a branched fragment thereof, or an oligomer thereof, or a monomer and/or macromer thereof. 
     
     
         79 . The method of  claim 65 , wherein the binding module specifically binds to a glycoprotein, carbohydrate, or both, specific to a particular blood antigen, type, group, or subgroup. 
     
     
         80 . The method of  claim 65 , wherein the binding module specifically binds to a polyaryletherketone (PAEK), a polyether ether ketone (PEEK), a polyethylene, a polypropylene, a polystyrene, a polytetrfluoroethylene, a polyvinylchloride, a polyamide, a para-aramid, a polyethylene terephthalate, a polyimide, a polycarbonate, a polypeptide, a polynucleotide, a glycoprotein, a protein, a phosphorylated protein, a modified protein, a lipid, a surfactant, lecithin, or a biosurfactant, or any combination thereof. 
     
     
         81 . The method of  claim 29 , said lignocellulosic polymer detection probe comprising a) a binding module that specifically binds to at least one lignocellulosic polymer and b) a reporter module that is spectroscopically detectable and wherein the binding module is a binding module polypeptide and the reporter module is a reporter module polypeptide that comprises a fluorescent protein and wherein the reporter module is not an antibody or a fragment thereof and wherein the binding module polypeptide is not an antibody or a fragment thereof; and
 measuring a property associated with the reporter module to determine the presence or absence of at least one lignocellulosic polymer in the biomass material based on specific binding of the probe to the at least one lignocellulosic polymer, wherein the fluorescent protein comprises Sirius, TagBFP, mTagBFP2, Azurite, EBFP2, mKalama1, Sirius, Sapphire, T-Sapphire, ECFP, mAmetrine, Cerulean, mCerulean3, SCFP3A, CyPet, mTurguoise, mTurquoise2, monomeric Midoriishi-Cyan, Aquamarine, eCFP, TagCFP, mTFP1, AmCyanl, EGFP, Emerald, Superfolder GFP, monomeric Azami Green, TurboGFP, TagGFP2, mUKG, mWasabi, Clover, mNeonGreen, eGFP, AcGFP1, ZGreen1, ZsYellow1, mBanana, EYFP, Topaz, Citrine, Venus, SYFP2, Ypet, IanRFP-deltaS83, mPapaya1, TagYFP, mOrange, mOrange2, monomeric Kusabira-Orange, mKOk, mKO2, mTangerine, mNectarine, TagRFP, TagRFP-T, mApple, mRuby, mRuby2, DsRed-Express2, DsRed-Express, tdTomato, DsRed-Monomer, DsRed-Monomer, DsRed2, AsRed2, mStrawberry, mCherry, HcRed1, FusionRed, mRaspberry, E2-Crimson, mPlum, HcRed-Tandem, mKate2, mNeptune, NirFP, TagRFP657, TagRFP675, iFP1.4, iRFP(iRFP713), iRFP670, iRFP682, iRFP702, iRFP720, iFP2.0, mIFP, mKeima Red, LSS-mKate1, LSS-mKate2, LSSmOrange, mBeRFP, PA-GFP, PATag RFP, Dendra2, Timer, PAmCherry, Kaede (green), Kaeda (red), KikGR1 (green), KikGR1 (red), PS-CFP2, mEos2 (green), mEos2 (red), mEos3.2 (green), mEos3.2(red), PSmOrange, Dropna, or any combination thereof and wherein the binding module is a carbohydrate-binding module (CBM) comprising CBM1, CBM2, CBM3, CBM3a CBM4, CBM5, CBM6, CBM9, CBM10, CBM11, CBM12, CBM14, CBM15, CBM17, CBM18, CBM19, CBM20, CBM21, CBM25, CBM27, CBM28, CBM33, CBM48, CBM49, or any combination thereof.