Choice of host cell line is essential for the functional glycosylation of the fragment crystallizable (Fc) region of human IgG1 inhibitors of influenza B viruses
Choice of host cell line is essential for the functional glycosylation of the fragment crystallizable (Fc) region of human IgG1 inhibitors of influenza B viruses
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DOI：
10.1101/719849
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发表时间：
2019-07
期刊：
影响因子：
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通讯作者：
P. Blundell;Dongli Lu;A. Dell;S. Haslam;R. Pleass
中科院分区：
文献类型：
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作者：
P. Blundell;Dongli Lu;A. Dell;S. Haslam;R. Pleass
抗体是携带与Fc连接的保守的N-连接的碳水化合物的糖蛋白，其存在和精细结构深刻地影响其体内免疫原性、药代动力学和功能属性。用于生产IgG的宿主细胞系对该糖基化具有重大影响，因为不同的系统表达不同的糖基化酶和转运蛋白，这有助于最终IgG-Fc糖基化谱的特异性和异质性。在此，我们比较了在HEK 293-F或CHO-K1系统中表达的两组聚糖适应性IgG 1-Fc突变体。我们发现，匹配的Fc突变体对之间的N-连接的聚糖的类型变化显着，特别是相对于唾液酸化。这些细胞系对糖基化的影响深刻地影响了工程化Fc与人或病原体受体相互作用的能力。例如，我们描述了Fc突变体，当在CHO-K1中表达而不是在HEK 293-F细胞中表达时，其有效地破坏乙型流感病毒介导的人红细胞凝集。
Antibodies are glycoproteins that carry a conserved N-linked carbohydrate attached to the Fc, whose presence and fine structure profoundly impacts on their in vivo immunogenicity, pharmacokinetics and functional attributes. The host cell line used to produce IgG has a major impact on this glycosylation, as different systems express different glycosylation enzymes and transporters that contribute to the specificity and heterogeneity of the final IgG-Fc glycosylation profile. Here we compare two panels of glycan-adapted IgG1-Fc mutants expressed in either the HEK 293-F or CHO-K1 systems. We show that the types of N-linked glycans between matched pairs of Fc mutants vary significantly, and in particular with respect to sialylation. These cell line effects on glycosylation profoundly influence the ability of the engineered Fcs to interact with either human or pathogen receptors. For example, we describe Fc mutants that potently disrupted influenza B-mediated agglutination of human erythrocytes when expressed in CHO-K1 but not in HEK 293-F cells.