Genetically Encoded, Multivalent Liquid Glycan Array (LiGA)

Autor: Nicholas J. Bennett, Mirat Sojitra, Ping Zhang, Susmita Sarkar, Chang-Chun Ling, Revathi Reddy, Corwin M. Nycholat, Michael R. Bell, Daniel Ferrer Vinals, Eric J. Carpenter, Emily Rodrigues, Ratmir Derda, Jasmine Maghera, Xiaochao Xue, Matthew S. Macauley, Ruixiang Blake Zheng, James C. Paulson, Shaurya Seth, Amira Khalil, Todd L. Lowary
Jazyk: angličtina
Rok vydání: 2020
Předmět:
DOI: 10.1101/2020.03.24.997536
Popis: 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.DedicationThe paper is dedicated to Laura L. Kiessling on the occasion of her 60th birthday.
Databáze: OpenAIRE