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.

中文翻译：

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DNA编码的聚糖图书馆作为研究聚糖-蛋白质相互作用的下一代工具

聚糖和聚糖结合蛋白（GBP）之间的相互作用介导了多种细胞功能，因此具有诊断和治疗意义。当前研究聚糖-GBP相互作用的领先策略需要专门的知识和仪器。在这项研究中，我们报告了一种研究聚糖-GBP相互作用的策略，该策略使用PCR，qPCR和下一代测序（NGS）技术，这些技术更易于使用。我们的耳机共轭编码连接（HCCL）策略将聚糖与独特的DNA编码结合在一起，这些DNA编码可在测序时指定聚糖部分和糖苷键。我们通过合成50种生物学上相关的聚糖（DEGL-50）的DNA编码的聚糖文库并探测针对25种靶蛋白（包括凝集素和抗体）的相互作用来证明该技术。数据显示溶液中的聚糖-GPB相互作用与含量较低，通量较低的ELISA分析一致。数据进一步表明，在使用较小的样本量的同时，如何使用单价或多价头戴式耳机来增加聚糖-GPB相互作用并丰富信号。我们的HCCL模块化策略的灵活性具有产生大型聚糖文库，促进高含量，高通量聚糖结合研究的潜力，并增加了获得低成本糖类分析的机会。