This fast-track STTR application proposes to synthesize structurally defined heparan sulfate and chondroitin sulfate oligosaccharides for biological research. Glycan Therapeutics is a biotech company specializing in synthesizing sulfated carbohydrates using innovative enzyme-based methods. This proposal is focused on the synthesis of large authentic oligosaccharides in the size range of 10-mers to 16-mers, as well as ultra-large click oligosaccharides in the size range of 30 saccharide residue long. A highly experienced research team, including Dr. Yongmei Xu (University of North Carolina), Dr. Robert Linhardt (Rensselaer Polytechnic Institute), Dr. Jian Liu (University of North Carolina) and Dr. Vijay Pagadala (Glycan Therapeutics), will carry out the study. Heparan sulfate and chondroitin sulfate are members of the glycosaminoglycan family present in large quantities on mammalian cell surfaces and in the extracellular matrix, displaying a wide range of biological functions. This proposal is designed to serve two purposes: (i) to develop a reliable and cost-effective method to synthesize structurally defined heparan sulfate and chondroitin sulfate oligosaccharides and structurally controlled polysaccharides to support glycoscience research; and (ii) to simplify this technology to allow nonglycoscientists to utilize this technology in a standard biochemistry laboratory setting. We anticipate that this proposal will provide valuable technical advances to glycoscience research.
Three specific aims are proposed:
Aim 1 is to synthesize a library of structurally defined heparan sulfate oligosaccharides with a size of up to hexadecasaccharides (16-mers);
Aim 2 is to develop novel chemoenzymatic methods to synthesize chondroitin sulfate oligosaccharides;
Aim 3 is to prepare structurally controlled click heparan sulfates and chondroitin sulfate oligosaccharides.
The purpose of this supplementary application is to request funding to purchase a bioreactor system. This instrument will be utilized to grow insect cells to produce both heparan sulfate and chondroitin sulfate biosynthetic enzymes for the enzymatic synthesis of both heparan sulfate and chondroitin sulfate oligosaccharides. The new bioreactor will increase our enzyme production capability by 5-10-fold, allowing us to complete the proposed synthesis.
