Natural, chimeric and hybrid glycosaminoglycan polymers and methods of making and using same

Abstract

The present invention relates to methodology for polymer grafting by a polysaccharide synthase and, more particularly, polymer grafting using the hyaluronate or chondroitin or heparin/heparosan synthases from Pasteurella multocida, in order to create a variety of glycosaminoglycan oligosaccharides having a natural or chimeric or hybrid sugar structure.

1 - 172 . (canceled)  
     
     
         173 . A method for producing a chimeric or hybrid glycosaminoglycan having a non-natural structure, comprising the steps of: 
 providing a functional acceptor, wherein the functional acceptor has at least two sugar units selected from the group consisting of uronic acid and hexosamine;    providing at least one glycosaminoglycan transferase, wherein the glycosaminoglycan transferase is selected from the group consisting of pmHAS, pmCS, pmHS, PglA, and mutants thereof; and    providing at least one of UDP-GlcUA, UDP-GlcNAc, UDP-Glc and UDP-GalNAc such that the at least one glycosaminoglycan transferase elongates the functional acceptor in a manner so as to provide a chimeric or hybrid glycosaminoglycan having a non-natural structure.    
     
     
         174 . The method of  claim 173 , wherein the functional acceptor is a plurality of functional acceptors immobilized on a substrate.  
     
     
         175 . The method of  claim 173 , wherein the functional acceptor is a plurality of functional acceptors in a liquid phase.  
     
     
         176 . A chimeric or hybrid glycosaminoglycan having a non-natural structure produced according to the methodology of  claim 173 , wherein the chimeric or hybrid glycosaminoglycan having a non-natural structure is composed of from about 1 to about 50 sugars.  
     
     
         177 . A library of natural glycosaminoglycan polymers produced according to the process comprising the steps of: 
 treating at least one functional acceptor with at least one glycosaminoglycan transferase in the presence of at least one UDP-sugar selected from the group consisting of UDP-GlcNAc, UDP-GlcUA, UDP-GalNAc, and UDP-GlcN.    
     
     
         178 . The product made according to the process of  claim 177 , wherein the functional acceptor is immobilized on a microtiter plate.  
     
     
         179 . The product made according to the process of  claim 177 , wherein the functional acceptor is immobilized on a microarray slide.  
     
     
         180 . The product made according to the process of  claim 177 , wherein the functional acceptor is in a liquid phase.  
     
     
         181 . The product made according to the process of  claim 180 , wherein the functional acceptor is in the liquid phase and is encoded with a distinct tag capable of decoding structure.  
     
     
         182 - 187 . (canceled)