Department of Chemistry, University of Alberta, Edmonton, AB, Canada.
Methods Mol Biol. 2024;2793:143-159. doi: 10.1007/978-1-0716-3798-2_10.
The M13 phage platform is a stable and monodisperse nanoscale carrier, which can be modified with different molecules by chemical conjugation strategies. Here, we describe M13 phage acylated on pVIII protein with a dibenzocyclooctyne reacting with azido glycan to yield 30-1500 copy numbers of glycan per phage and monitored by MALDI-TOF spectrometry to generate multivalent glycoconjugates that contain desired densities of glycans. We prepared the liquid glycan arrays (LiGA) such that both the structure and density of glycans were encoded in the DNA of the bacteriophage. The LiGA can be used to validate the binding properties of glycans to purified lectins and explore the effect of glycan density on such binding. From a mixture of multivalent glycan probes, LiGAs can also identify the glycoconjugates with optimal avidity necessary for binding to lectins on living cells in vitro and live animals in vivo.
M13 噬菌体平台是一种稳定且单分散的纳米载体，可通过化学偶联策略用不同的分子进行修饰。在这里，我们描述了用二苯并环辛炔修饰 pVIII 蛋白的 M13 噬菌体与叠氮聚糖反应，得到每个噬菌体带有 30-1500 个聚糖拷贝，并通过 MALDI-TOF 光谱法监测，生成具有所需聚糖密度的多价糖缀合物。我们制备了液体糖阵列 (LiGA)，使得聚糖的结构和密度都编码在噬菌体的 DNA 中。LiGA 可用于验证聚糖与纯化的凝集素的结合特性，并研究聚糖密度对这种结合的影响。从多价糖探针的混合物中，LiGA 还可以鉴定出与体外培养的活细胞和体内活动物上的凝集素结合所需的最佳亲和力的糖缀合物。
