DNA Encoded Glycan Libraries as a next-generation tool for the study of glycan-protein interactions

Kondengaden, S. M.; Zhang, J.; Zhang, H.; Parameswaran, A.; Kondengadan, S. M.; Pawar, S.; Puthengot, A.; Sunderraman, R.; Song, J.; Polizzi, S. J.; Wen, L.; Wang, P. G.

doi:10.1101/2020.03.30.017012

bioRxiv · 10.1101/2020.03.30.017012

DNA Encoded Glycan Libraries as a next-generation tool for the study of glycan-protein interactions

Kondengaden, S. M.·Zhang, J.·Zhang, H.·Parameswaran, A.·Kondengadan, S. M.·Pawar, S.·Puthengot, A.·Sunderraman, R.·Song, J.·Polizzi, S. J.·Wen, L.·Wang, P. G.

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Abstract

Interactions between glycans and glycan-binding proteins (GBPs) mediate diverse cellular functions, and therefore are of diagnostic and therapeutic significance. Current leading strategies for studying glycan-GBP interactions require specialized knowledge and instrumentation. In this study, we report a strategy for studying glycan-GBP interactions that uses PCR, qPCR and next-generation sequencing (NGS) technologies that are more routinely accessible. Our headpiece conjugation-code ligation (HCCL) strategy couples glycans with unique DNA codes that specify glycan sugar moieties and glycosidic linkages when sequenced. We demonstrate the technology by synthesizing a DNA encoded glycan library of 50 biologically relevant glycans (DEGL-50) and probing interactions against 25 target proteins including lectins and antibodies. Data show glycan-GPB interactions in solution that are consistent with lower content, lower throughput ELISA assays. Data further demonstrate how monovalent and multivalent headpieces can be used to increase glycan-GPB interactions and enrich signals while using smaller sample sizes. The flexibility of our modular HCCL strategy has potential for producing large glycan libraries, facilitating high content-high throughput glycan binding studies, and increasing access to lower cost glyco-analyses. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=60 SRC="FIGDIR/small/017012v1_ufig1.gif" ALT="Figure 1"> View larger version (17K): org.highwire.dtl.DTLVardef@fb1c1dorg.highwire.dtl.DTLVardef@1f2c3ceorg.highwire.dtl.DTLVardef@11546e7org.highwire.dtl.DTLVardef@1e21a83_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Kondengaden, S. M., Zhang, J., Zhang, H., Parameswaran, A., Kondengadan, S. M., Pawar, S., Puthengot, A., Sunderraman, R., Song, J., Polizzi, S. J., Wen, L., Wang, P. G.. 2020-03-31. DNA Encoded Glycan Libraries as a next-generation tool for the study of glycan-protein interactions. https://doi.org/10.1101/2020.03.30.017012

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