Repeat sequence protein polymer active agent congjugates, methods and uses

Abstract

Biomolecular conjugates are provided which comprise the conjugation product of a repeat sequence protein polymer and at least one active agent. Additional aspects provide methods for their manufacture and various industrial and consumer applications.

What is claimed is:  
     
         1 . Biomolecular conjugates comprising the conjugation product of a repeat sequence protein polymer and at least one active agent.  
     
     
         2 . The biomolecular conjugates as recited in  claim 1 , wherein the repeat sequence protein polymer comprises a repeating amino acid sequence unit derived from elastin, collagen, abductin, byssus, flagelliform silk, dragline silk, gluten high molecular weight subunit, titin, fibronectin, leminin, gliadin, glue polypolypeptide, ice nucleating protein, keratin, mucin, RNA polymerase II, resalin or a mixture thereof.  
     
     
         3 . The biomolecular conjugates as recited in  claim 1 , wherein the repeat sequence protein polymer formula comprises:  
       T y [(A n ) x (B) b (A′ n′ ) x′ (B′) b′ (A″ n ″) x ″] i T′ y′   
       wherein: 
 T and T′ each comprise an amino acid sequence of from about 1 to about 100 amino acids, wherein the amino acid sequence of T′ is the same as or different from the amino acid sequence of T;  
 y and y′ are each an integer from 0 to 1, wherein the integer of y′ is the same as or different from the integer of y;  
 A, A′ and A″ are each individual repeating sequence units comprising from about 3 to about 30 amino acids, wherein the amino acid sequence of A′ and the amino acid sequence of A″ are the same as or different from the amino acid sequence of A;  
 n, n′, and n″ are integers of at least 2 and not more than 250;  
 x, x′ and x″ are each 0 or an integer of at least 1, wherein each integer varies to provide for at least 30 amino acids in the A′, A′ and A″ individual repeating sequence units, and wherein the integer of x′ and the integer of x″ are the same as or different from the integer of x;  
 B and B′ each comprise an amino acid sequence of from about 4 to about 50 amino acids, wherein the amino sequence of B′ is the same as or different from the amino acid sequence of B;  
 b and b′ are each an integer from 0 to 3, wherein the integer of b′ is the same as or different from the integer of b;  
 i is an integer from 1 to 100.  
 
     
     
         4 . The biomolecular conjugates as recited in  claim 3 , wherein T and T′ comprise an amino acid sequence from about 1 to about 60 amino acids.  
     
     
         5 . The biomolecular conjugates as recited in  claim 3 , wherein T and T′ comprise an amino acid sequence with fewer than 20% of the total number of amino acids in the repeat sequence protein polymer.  
     
     
         6 . The biomolecular conjugates as recited in  claim 3 , wherein B comprises an amino acid sequence with a biological or chemical function or activity.  
     
     
         7 . The biomolecular conjugates as recited in  claim 3 , wherein B′ comprises an amino acid sequence with a biological or chemical function or activity.  
     
     
         8 . The biomolecular conjugates as recited in  claim 3 , wherein i is an integer from 1 to 50.  
     
     
         9 . The biomolecular conjugates as recited in  claim 3 , wherein i is an integer from 1 to about 30.  
     
     
         10 . The biomolecular conjugates as recited in  claim 3 , wherein A, A′, and A″ comprise SEQ ID NO. 1, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 8, SEQ ID NO. 9, SEQ ID NO. 10, SEQ ID NO. 11, SEQ ID NO. 12, SEQ ID NO. 13, SEQ ID NO. 14, SEQ ID NO. 15, SEQ ID NO. 16, SEQ ID NO. 17, SEQ ID NO. 18, SEQ ID NO. 20 or a combination thereof.  
     
     
         11 . The biomolecular conjugates as recited in  claim 3 , wherein A, A′, and A″ comprise SEQ ID NO. 1, SEQ ID NO. 3 or a combination thereof.  
     
     
         12 . The biomolecular conjugates as recited in  claim 1 , wherein the repeat sequence protein polymer comprises SEQ ID NO. 19.  
     
     
         13 . The biomolecular conjugates as recited in  claim 1 , wherein the at least one active agent is selected from the group consisting of: silicones, fragrances, dyes, tints, UV actives, sunscreens, lanolin, vitamins, bleaches, thickening agents, proteins, peptides, enzymes, antimicrobials, preservatives, and combinations thereof.  
     
     
         14 . The biomolecular conjugates as recited in  claim 13 , wherein the at least one active agent comprises a silicone.  
     
     
         15 . The biomolecular conjugates as recited in  claim 14 , wherein the silicone is selected from the group consisting of: silicones comprising an epoxy functional group, polydimethyl siloxanes, silicone resins, silicone surfactants, alkyl modified silicones, gum-silocone fluid blends, phenyl silicone fluids, dimethyl silicone fluids, quaternized ammonium salts of suitable silicones, silicones comprising one or more amino functional groups, and combinations thereof.  
     
     
         16 . A personal care composition comprising the biomolecular conjugates as recited in  claim 1 .  
     
     
         17 . The personal care composition as recited in  claim 16  comprising a hair care composition, a skin care composition, a nail care composition, a cosmetic composition, an oral care composition, or an over-the-counter pharmaceutical composition.  
     
     
         18 . The personal care composition as recited in  claim 16  wherein the biomolecular conjugates comprise from about 0.001% to about 10% by weight of the composition  
     
     
         19 . The personal care composition as recited in  claim 16  wherein the biomolecular conjugates comprise from about 0.01% to about 5% by weight of the composition.  
     
     
         20 . The personal care composition as recited in  claim 16  wherein the biomolecular conjugates comprise from about 0.01% to about 1% by weight of the composition  
     
     
         21 . A process for making a personal care composition comprising combining a biomolecular conjugate with an acceptable carrier or excipient to obtain a personal care composition, wherein the biomolecular conjugate comprises a conjugation product of a repeat sequence protein polymer and at least one active agent.  
     
     
         22 . The biomolecular conjugates as recited in  claim 1  formulated into an emulsion comprising, by weight of the emulsion composition:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                     
                 
                     
                   water 
                   qs 
                 
                     
                   emulsifier(s) 
                      1-5%  
                 
                     
                   thickener(s)/stabilizers 
                    0.1-3%  
                 
                     
                   emollient(s) 
                      2-10% 
                 
                     
                   opacifier(s) 
                      0-10% 
                 
                     
                   humectant(s) 
                      0-10% 
                 
                     
                   system 
                   0.001-10% 
                 
                     
                   functional ingredients 
                   0.001-25% 
                 
                     
                   preservative(s) 
                   qs 
                 
                     
                   finishing ingredients 
                   qs 
                 
                     
                     
                 
                     
                     
                 
             
                
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         23 . A personal care product composition comprising the emulsion as recited in  claim 22 .  
     
     
         24 . The biomolecular conjugates as recited in  claim 1  formulated into a surfactant system comprising, by weight of the surfactant system composition:  
       
         
           
                 
                 
                 
               
                     
                     
                 
                     
                     
                 
                     
                   water 
                   qs 
                 
                     
                   primary surfactant(s) 
                    0.1-15% 
                 
                     
                   secondary surfactant(s) 
                    0.1-10% 
                 
                     
                   rheology modifier(s) 
                    0.1-5%  
                 
                     
                   alcohol(s) 
                      0-25% 
                 
                     
                   system(s) 
                   0.001-10% 
                 
                     
                   functional ingredient(s) 
                      0-10% 
                 
                     
                   conditioning ingredient(s) 
                      0-5%  
                 
                     
                   preservative(s) 
                   qs 
                 
                     
                   finishing ingredient(s) 
                   qs 
                 
                     
                     
                 
                     
                     
                 
             
                
                
               
               
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         25 . A personal care product composition comprising the surfactant system as recited in  claim 24 .  
     
     
         26 . The biomolecular conjugate as recited in  claim 1  wherein the repeat sequence protein polymer comprises a silk elastin polymer and the at least one active agent comprises a protein or peptide, and further wherein the conjugation product comprises a fusion protein.  
     
     
         27 . The biomolecular conjugate as recited in  claim 26  wherein the silk elastin polymer comprises SELP47K (SEQ. ID. NO. 19), and the protein or peptide comprises any protein or peptide suitable for a desired application and which is capable of forming a conjugation product comprising a fusion protein with SELP47K (SEQ. ID. NO. 19).  
     
     
         28 . The biomolecular conjugate as recited in  claim 26 , wherein the peptide comprises an antimicrobial peptide.  
     
     
         29 . The biomolecular conjugate as recited in  claim 26  comprising CAM-SELP47K (SEQ. ID. NO. 32).  
     
     
         30 . The biomolecular conjugate as recited in  claim 26  comprising MBI-SELP47K (SEQ. ID. NO. 33)  
     
     
         31 . The biomolecular conjugate as recited in  claim 26 , wherein the protein comprises a green fluorescent protein.  
     
     
         32 . The biomolecular conjugate as recited in  claim 26  comprising GFP-SELP47K (SEQ. ID. NO. 34).  
     
     
         33 . The biomolecular conjugate as recited in  claim 26 , wherein the peptide comprises a cotton binding peptide.  
     
     
         34 . The biomolecular conjugate as recited in  claim 26  comprising CBP-SELP47K-S2E1 (SEQ. ID. NO. 35).  
     
     
         35 . The biomolecular conjugate as recited in  claim 26 , wherein the peptide comprises P4.  
     
     
         36  The biomolecular conjugate as recited in  claim 26  comprising P4-SELP47K (SEQ. ID. NO. 36).  
     
     
         37 . The biomolecular conjugate as recited in  claim 26  comprising at least two active agents, and wherein the at least two active agents are different.  
     
     
         38 . A method of producing a biomolecular conjugate comprising a fusion protein conjugate comprising a conjugation product of a repeat sequence protein polymer and at least one active agent comprising a protein or peptide, the method comprising: selecting the repeat sequence protein polymer and the protein or peptide active suitable for a desired application; obtaining a gene encoding the repeat sequence protein polymer and a gene encoding the at least one active agent comprising a protein or peptide; constructing a conjugate gene from the gene encoding the repeat sequence protein polymer and the gene encoding the at least one active agent comprising a protein or peptide; expressing the conjugate gene to form an expression product comprising the fusion protein conjugate; fermenting the expression product comprising the fusion protein conjugate; and purifying the fusion protein conjugate.  
     
     
         39 . The method as recited in  claim 38  comprising at least two active agents.  
     
     
         40 . The method as recited in  claim 38  wherein the at least one active agent comprises a protein or a peptide comprising an antimicrobial peptide, P4, a cotton binding peptide, green fluorescent protein, or a combination thereof.  
     
     
         41 . A method of providing biomaterial adapted for at least one predetermined desirable function comprising: selecting a biomolecular conjugate comprising a conjugation product of a repeat sequence protein polymer and at least one active agent, wherein the repeat sequence protein polymer comprises a silk elastin polymer and the at least one active agent comprises a protein or peptide, and further wherein the conjugation product comprises a fusion protein, according to the predetermined desirable function; and incorporating the biomolecular conjugate into a material.  
     
     
         42 . The method as recited in  claim 41 , wherein the method provides biomaterial adapted for more than one predetermined desirable function and wherein the biomolecular conjugate comprises at least two active agents according to the predetermined desirable function.  
     
     
         43 . A biomaterial adapted for at least one predetermined desirable function comprising at least one biomolecular conjugate comprising a conjugation product of a repeat sequence protein polymer and at least one active agent, wherein the repeat sequence protein polymer comprises a silk elastin polymer and the active agent comprises a protein or peptide, and further wherein the conjugation product comprises a fusion protein.  
     
     
         44 . The biomaterial as recited in  claim 43  wherein the at least one biomolecular conjugate comprises AMP-SELP47K, GFP-SELP47K, P4-SELP47K, CBP-SELP47K, or some combination thereof.  
     
     
         45 . The biomaterial as recited in  claim 44 , wherein the biomolecular conjugate comprises AMP-SELP47K, and wherein the biomaterial comprises a textile, a filter, a cream or a coating.  
     
     
         46 . The biomaterial as recited in  claim 45 , wherein the textile comprises a protective garment.  
     
     
         47 . The biomaterial as recited in  claim 46  wherein the protective garment comprises a surgical gown or a garment designed for biological warfare defense.  
     
     
         48 . The biomaterial as recited in  claim 43 , wherein the biomolecular conjugate comprises GFP-SELP47K, and wherein the biomaterial elicits a detectable output in response to a predetermined stimuli.  
     
     
         49 . The biomaterial as recited in  claim 48 , wherein the biomaterial comprises a genetics research tool or a search and/or identification tool.  
     
     
         50 . The biomaterial as recited in  claim 43 , wherein the biomolecular conjugate comprises P4-SELP47K, wherein the biomaterial comprises a textile, dressing, cream or coating for the protection of ultraviolet radiation sensitive surfaces.  
     
     
         51 . A repeat sequence protein polymer comprising the biomolecular conjugate as recited in  claim 1.