Natural Glycans for Functional Glycomics

Project Summary Abstract Progress in glycomicsthe systematic study of the structure and function of glycanslags far behind that of genomics and proteomics due to the more complex structures of glycanswhose biosynthesis is posttranslational and not template drivenA major limitation in glycomics is the absence of powerful amplification technologies such as PCRrecombinant technology and automated chemical synthesiswhich limits the availability of biologically relevant glycans for functional studiesTo address this limitationwe developed a novel technologyoxidative release of natural glycansORNGwhich uses simple oxidation by sodium hypochloritehousehold bleachto degrade peptides and nucleic acids and release bleach resistant glycans from the glycoconjugates of natural sources such as animal plant tissuesWe have demonstrated that unlike current methods for releasing glycansORNG is a scalable process which has allowed us to generate unprecedented amounts of glycans from up tokg of animal tissueThusthe absence of glycan amplification technology can be addressed by increasing the amount of starting materialwhich results in large quantities of releasedbiologically relevant glycansGlycan vendors manufacture N glycans using nonscalable methods resulting in limited amounts of material and high costsg packages for $to $Our scalable process will allow us to generate gram quantities of high mannosepaucimannoseand complextype N glycansand the economy realized by the large scale will allow us to provide mg quantities of N glycans at andltof current costs making these glycans available to non specialized laboratories in mg quantities for functional studiesWe have demonstrated the utility of our process for manufacturing mg quantities of Nglycanswhich are not currently available in these quantitiesHoweverORNG is unique in that it releases all classes of glycans in a singlerapid reactionand we have currently optimized it for release of N glycans to demonstrate feasibilityHerewe are requesting funds to develop a streamlineddownstream processfor purification of all classes of glycans including glycosaminoglycansO linked glycans and glycosphingolipidlinked glycans from a single oxidation runVendors of O linked glycans and glycosphingolipid linked glycans generally manufacture them by chemical or chemo enzymatic synthesiswhich is expensive and limited to single products that are selected based on our limited information of what structures exist in natureSince ORNG uses kg quantities of natural products as starting materialthe resulting glycans will represent a combinatorial library of the source natural product containing not only the knowncommon glycansbut also minor glycans that have not been previously detectedThis project will result in a robust manufacturing process for purifiednatural NOGSL linked glycans that we can offer at affordable pricesIn additionwe will manufacture the corresponding shotgun glycan microarrays based on thesecombinatoriallibraries that can be interrogated with biologically relevant GBPs to discover novel glycans with unique functionsNarrative Progress in glycomicsthe systematic study of the structure and function of glycanslags far behind that of genomics and proteomics due to lack of purifiedbiologically relevant glycans at quantities required for functional studiesUsing its proprietary process for oxidative release of natural glycansORNGNatGlycanLLC has developed a commercialscalable process to generate unprecedented quantities of all classes of glycans from natural sources in a singlerapid reaction which addresses the lack of amplification technology in this areaHerewe propose to develop a streamlineddownstream processthat will make naturalbiologically relevant glycans available to the biomedical community at affordable prices based on economy of scale