Genetically Encoded, Multivalent Liquid Glycan Array (LiGA)

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...

Full description

Saved in:
Bibliographic Details
Published inNature chemical biology Vol. 17; no. 7; pp. 806 - 816
Main Authors Sojitra, Mirat, Sarkar, Susmita, Maghera, Jasmine, Rodrigues, Emily, Carpenter, Eric, Seth, Shaurya, Vinals, Daniel Ferrer, Bennett, Nicholas, Reddy, Revathi, Khalil, Amira, Xue, Xiaochao, Bell, Michael, Zheng, Ruixiang Blake, Zhang, Ping, Nycholat, Corwin, Bailey, Justin J., Ling, Chang-Chun, Lowary, Todd L., Paulson, James C., Macauley, Matthew S., Derda, Ratmir
Format Journal Article
LanguageEnglish
Published 06.05.2021
Online AccessGet full text

Cover

More Information
Summary: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.
Bibliography:N.J.B. constructed and characterized all LiGA plasmids and libraries, M.S., S.Sa., J.M., D.F.V., R.R., performed isolation of phage clones, modifications of phage by glycans, MALDI analysis, protein-binding experiments, and in vitro assays. M.S., E.R., J.M., and R.D. performed cell-based assays, M.S.M and S.Sa. performed animal experiments, E.J.C., M.S. and R.D. performed statistical analysis., S.Se., performed custom analysis of public CFG data., T.L.L., P.Z., C.C.L., C.N. and J.C.P. contributed synthetic glycan reagents, A.K. and M.B. constructed synthetic antigens to raise and characterize anti-galactofuranose antibodies, which were produced by S.Sa. X.X. and R.B.Z. constructed antigens for incorporation into the LiGA. R.D., and M.S. wrote the manuscript, R.D., M.S.M., T.L.L., and J.C.P. edited the final manuscript and contributed intellectual and strategic input. All authors approved the final manuscript.
Contributions
present address: Pharmaceutical Chemistry Department, Faculty of Pharmacy, British University in Egypt, El-Sherouk city, Cairo 11837, Egypt.
ISSN:1552-4450
1552-4469
DOI:10.1038/s41589-021-00788-5