Abstract
The Central Dogma of Biology does not allow for the study of glycans using DNA sequencing. We report a “Liquid Glycan Array” (LiGA) platform comprising a library of DNA ‘barcoded’ M13 virions that display 30-1500 copies of glycans per phage. A LiGA is synthesized by acylation of phage pVIII protein with a dibenzocyclooctyne, followed by ligation of azido-modified glycans. Pulldown of the LiGA with lectins followed by deep sequencing of the barcodes in the bound phage decodes the optimal structure and density of the recognized glycans. The LiGA is target agnostic and can measure the glycan-binding profile of lectins such as CD22 on cells in vitro and immune cells in a live mouse. From a mixture of multivalent glycan probes, LiGAs identifies the glycoconjugates with optimal avidity necessary for binding to lectins on living cells in vitro and in vivo; measurements that cannot be performed with canonical glass slide-based glycan arrays.
Dedication The paper is dedicated to Laura L. Kiessling on the occasion of her 60th birthday.
Footnotes
Two authors have been added to the author list. Summary of the synthesis of the glycan components was added to the Supporting Table S2. Explicit p-values have been added to Figures 2 and 6 of the main text instead of * designations. Clarification was added to Figure legends of Main Text Figures 2-5 and S7-S11 Supporting Figures on statistical methods. Explicit MALDI data and PFU titer data was included in the Data.zip folder. Minor typow have been corrected in data files. Additional references have been added to the main text and supporting information. Minor typos have been corrected in the Main Text, Acknowledgement, Contributions, and supporting Information.
