Monoclonal antibodies specific for biologically relevant glycans

PROJECT SUMMARY We are combining two innovative technologiesNovAbandapos s Variable Lymphocyte ReceptorVLRmonoclonal antibody technology and NatGlycanandapos s Oxidative Release of Natural GlycansORNGtechnologyto create a high throughputhigh capacity process for discovery of a new class of binding reagents for biologically relevant glycansi ethe glycans of tissuescells and glycoproteinsSuch reagents are essential to detectinglocalizingquantifyingand establishing biological function sof specific glycans in normal and disease cells and tissuesand may additionally lead to development of new diagnostic and therapeutic modalitiesThe absence of such reagents is considered a significant barrier to understanding the role of glycans in health and diseaseVLRs are the antigen receptors of the adaptive immune system of jawless vertebrateslamprey and hagfishare composed of tandemly arranged highly diverse leucine rich repeatLRRstructural motifs with diversity comparable to the human immunoglobulinIgantibody repertoireand possess a distinctive more rigidsheet binding site geometry vs Ig antibodies that is entirely contained within a single polypeptide structureVertebrates split into jawless and jawed vertebrate lineages approximatelymillion years agoThe evolutionary separation of jawless vertebrates from humans and other mammalsand the distinctive binding site geometry of VLRs potentially provide novel specificities not accessed with Ig antibodiesThis expectation is supported by the discovery of VLRs that bind glycans with affinity and specificity superior to that typically achieved with Ig antibodies and indicate that VLRs are uniquely wellsuited to binding and discriminating glycan structuresORNG is a technology for releasing glycans from natural sourcese gtissuescells and glycoproteinspurifying the released glycansdetermining structures of the purified glycansand chemically modifying glycans for attachment to solid surfacesIn combining these technologies biological sourcese ghuman cells or glycoproteinsare utilized to immunize lamprey and these same biological sources utilized to derive glycans via NatGlycanandapos s ORNG technology with which to select screen for lamprey VLR monoclonal antibodies that specifically bind each of the purified glycansWe expect this process to yield dozens of monoclonal VLRseach with a distinct glycan binding specificity from each lamprey immunization regimen with cellular and glycoprotein immunogensThe VLR ORNG technology is therefore potentially a transformative for the rapidhigh throughput production of specific binding reagents for the diverse glycan structures that comprise the human glycomePROJECT NARRATIVE Glycans participate in cell adhesion and traffickingprotein foldingstabilityand localizationmembrane integrityand cell signalingGlycans regulate both innate and adaptive immunityare frequently aberrant in cancerous cells and promote tumor progressionand are implicated in protection against neurodegenerative diseaseThrough this application we will combine two innovative technologies to create a high throughputhigh capacity capability for producing glycan binding reagents with which to detectlocalizequantify and modulate function sof glycans to facilitate a better understanding of the role of glycans in health and disease