Danya Medina-Carrasco, Lisandra García de Castro Cuspineda, Luis Gabriel González-Lodeiro, Satomy Pousa, Miladys Limonta, Vivian Huerta Galindo
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引用次数: 0
摘要
重组糖蛋白的特性可通过添加用于异源表达的细胞所特有的寡糖结构而改变。我们采用了一种方法来获得糖肽制剂,这种方法有助于阐明碳水化合物在糖蛋白的免疫原性和抗原性中的作用。该方法包括消化蛋白质,然后选择性地捕获与二肽/三肽结合的寡糖,并通过化学交联将其接枝到非糖基化受体蛋白上。从 C-RBD-H6 PP 蛋白(Abdala 疫苗的有效成分)中提取的糖肽有效地接枝到了非糖基化蛋白上,这一点已通过 Western 印迹得到证明。
Properties of recombinant glycoproteins can be altered by the addition of oligosaccharide structures specific to the cells used for its heterologous expression. A methodology was implemented to obtain a glycopeptide preparation useful to elucidate the role of carbohydrates in the immunogenicity and antigenicity of glycoproteins. It consists on the digestion of the protein, followed by selective capture of the oligosaccharides bound to di-/tripeptides, and their grafting onto a non-glycosylated receptor protein by chemical crosslinking. Glycopeptides derived from C-RBD-H6 PP protein, the active ingredient of the Abdala vaccine were efficiently grafted onto a non-glycosylated protein as evidenced by western blotting.