Glycocalyx Scaffolding to Control Cell Surface Glycan Displays.
Glycocalyx Scaffolding to Control Cell Surface Glycan Displays.
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DOI：
10.1002/cpch.40
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发表时间：
2018-06
影响因子：
--
通讯作者：
Godula K
中科院分区：
文献类型：
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作者：
Huang ML;Tota EM;Verespy S 3rd;Godula K
该单元描述了一种用合成糖蛋白和糖脂模拟物重塑细胞的方案，这些糖脂模拟物用脂质锚定物官能化，允许细胞表面显示预定义纳米级排列的特定聚糖结构。将CHO-Lec 2细胞制备为贴壁单层，并在达到汇合后与糖材料一起孵育。含有α 2，3-唾液酸乳糖聚糖的合成糖共聚物用于以远离细胞表面突出的3D多价配体的形式装饰细胞表面，而α 2，6-唾液酸乳糖糖脂缀合物用于将动态2D阵列中的聚糖锚在细胞膜附近。洗涤后，可以使用对α 2，3-或α 2，6-连接的唾液酸具有特异性的凝集素分析模拟掺入和聚糖展示。流式细胞术数据显示，用任一糖缀合物模拟物的细胞表面重塑以剂量依赖性方式有效地发生。也可以同时使用糖缀合物的组合以产生具有可调组成和组织的混合糖萼。
This unit describes a protocol for remodeling cells with synthetic glycoprotein and glycolipid mimetics that are functionalized with lipid anchors, allowing for cell surface display of specific glycan structures in predefined nanoscale arrangements. CHO-Lec2 cells are prepared as adherent monolayers and, after reaching confluence, are incubated with the glycomaterials. Synthetic glycopolymers bearing α2,3-sialyllactose glycans are used to decorate cellular surfaces in the form of 3D multivalent ligands projecting away from the cell surface, while α 2,6-sialyllactose glycolipid conjugates are used to anchor glycans in dynamic 2D arrays proximal to the cell membrane. Following washing, mimetic incorporation and glycan display can be analyzed using lectins with specificity for α 2,3- or α 2,6-linked sialic acids. Flow cytometry data reveals that cell surface remodeling with either glycoconjugate mimetic occurs efficiently in a dose-dependent manner. Combinations of glycoconjugates can also be employed simultaneously to generate a mixed glycocalyx with tunable composition and organization.