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单克隆抗体糖基化与糖基化工程抗体 (上)

自从人们发现单克隆抗体(MAbs)的应用价值后,它已被广泛的应用于科学研究及临床疾病的治疗中。目前MAbs主要被应用在癌症,自身免疫性疾病,感染性疾病及炎症的治疗。

MAbs的各种作用模式包括对抗原的中和作用,补体依赖的细胞毒作用(CDC)以及抗体依赖的细胞毒作用(ADCC)。抗体的中和作用依赖于Fab区与抗原上的表位相结合,从而阻止因生物分子相互作用而引起的疾病进展。通过CDC和ADCC作用的MAbs(所谓的“Fc效应功能”)依赖于两阶段的作用:第一阶段:抗体Fab区与抗原的表位结合(例如:癌细胞表面上的CD20受体)。第二阶段,各种类型的免疫细胞可以结合到抗体的Fc区,从而引发免疫系统中“效应”细胞的募集。这些免疫细胞分泌一些细胞因子可最终导致抗体结合细胞的死亡。CDC和ADCC涉及到不同的受体、效应细胞和靶细胞死亡模式,但是均受到抗体Fc功能区糖基化类型的显著影响。如下图中,抗体Fc段重链第297位天冬酰胺残基处为Fc段糖基化位点,此处糖基化形式可影响Fc段肽链的空间结构进而影响Fc末端与效应细胞的结合。

1

抗体Fc段Asn297位点的聚糖是由乙酰氨基葡糖,甘露糖,半乳糖,岩藻糖, N-羟乙酰基神经氨酸,N-乙酰神经氨酸等多种单糖组成的聚糖链。这些单糖通过不同的搭配组成多聚糖。因此,通过细胞发酵产生的抗体其实是具有相同氨基酸序列及不同聚糖链抗体的混合物。通过CHO细胞,NS0细胞发酵表达的抗体常见Fc段糖基化形式主要有以下几种:

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目前上市的抗体药物大多数是由CHO细胞及NS0细胞表达生产的,由于这两种细胞均为鼠源细胞,因此其表达的抗体多种糖基化形式对于人来说是外源性的。因而这些抗体分子在体内会产生免疫原性。利用基因编辑技术改变抗体表达细胞的糖基化相关基因,可产生活性更高免疫原性更低的糖基化工程抗体。在下篇中小编将会向大家介绍糖基化工程抗体一些研究及应用情况。

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