RT Journal Article
SR Electronic
T1 DNA Encoded Glycan Libraries as a next-generation tool for the study of glycan-protein interactions
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.03.30.017012
DO 10.1101/2020.03.30.017012
A1 Shukkoor M. Kondengaden
A1 Jiabin Zhang
A1 Huajie Zhang
A1 Aishwarya Parameswaran
A1 Shameer M. Kondengadan
A1 Shrikant Pawar
A1 Akhila Puthengot
A1 Rajshekhar Sunderraman
A1 Jing Song
A1 Samuel J. Polizzi
A1 Liuqing Wen
A1 Peng George Wang
YR 2020
UL http://biorxiv.org/content/early/2020/03/31/2020.03.30.017012.abstract
AB 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.