Developing high-affinity nanobodies to tumor-associated glycan antigens

Project SummaryGlycans are known to be involved in including cell growth and differentiationcell cell communicationimmune responseand intracellular signaling eventsGlycans also feature prominently in human diseaseIn many different human disordersthe cellular glycome is alteredThe key unsolved problems in the glycosciences are the tools for study the temporal and spatial distribution of glycans on in intact cellswhich is important to the revelation of glycan related biological eventsWe propose to develop a robustintegrated platform for the rapid discovery and characterization of high affinity nanobodiesi esingle domain antibodysdAbsagainst defined glycan structures of biomedical importanceThe proposed project seeks to overcome the critical barriers in producing high affinity glycan specific nanobodiesipoor immunogenicity of glycans iidifficulties and labor intensive nature for immobilization of carbohydrate antigens onto carrier as immunogensIn this Phase I proof of concept demonstrationwe will select and validate high affinity nanobodies to tumor associated glycans antigens of Thomsen Friedenreich antigenTnGal bGalNActhe sialyl TnNeu AcGalNAcantigenand sialyl LewisxNeu AcGalFucGlcNAcThe long term goal is to buildup libraries of nanobodies to a diversity of glycan structuresSuch an effort is anticipated to yield a set of well characterized new glycan specific affinity reagents that are suitable for probing temporal and spatial expression of glycans in cells and tissuesand provide accessible reagents for `non glycoandaposexperts in biology and biomedical field Project NarrativeTo unravel the role that glycans play in both health and diseasenew tools are needed to study the temporal and spatial distribution of glycans on in intact cellsThis proposed research is to develop a robustintegrated platform for the rapid discovery and characterization of high affinity nanobodies against defined glycan structures of biomedical importanceDue to the unique properties of nanobodiessmall sizehigh stabilityand tissue penetrationthe glycan recognizing nanobodies will be suitable for probing the temporal and spatial expression of glycans in cells and tissues