具有三种不同N-连接Fc寡糖(高甘露糖型、杂合型和复合型)的非岩藻糖基化治疗性IgG1抗体之间的生物学活性比较。

Comparison of biological activity among nonfucosylated therapeutic IgG1 antibodies with three different N-linked Fc oligosaccharides: the high-mannose, hybrid, and complex types.

作者信息

Kanda Yutaka, Yamada Tsuyoshi, Mori Katsuhiro, Okazaki Akira, Inoue Miho, Kitajima-Miyama Kazuko, Kuni-Kamochi Reiko, Nakano Ryosuke, Yano Keiichi, Kakita Shingo, Shitara Kenya, Satoh Mitsuo

机构信息

Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd, 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan.

出版信息

Glycobiology. 2007 Jan;17(1):104-18. doi: 10.1093/glycob/cwl057. Epub 2006 Sep 29.

Abstract

The structure of asparagine-linked oligosaccharides attached to the antibody constant region (Fc) of human immunoglobulin G1 (IgG1) has been shown to affect the pharmacokinetics and antibody effector functions of antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). However, it is still unclear how differences in the N-linked oligosaccharide structures impact the biological activities of antibodies, especially those lacking core fucose. Here, we succeeded in generating core fucose-lacking human IgG1 antibodies with three different N-linked Fc oligosaccharides, namely, a high-mannose, hybrid, and complex type, using the same producing clone, and compared their activities. Cultivation of an alpha-1,6-fucosyltransferase (FUT8) knockout Chinese hamster ovary cell line in the presence or absence of a glycosidase inhibitor (either swainsonine or kifunensine) yielded antibody production of each of the three types without contamination by the others. Two of three types of nonnaturally occurring atypical oligosaccharide IgG1, except the complex type, reduced the affinity for both human lymphocyte receptor IIIa (FcgammaRIIIa) and the C1q component of the complement, resulting in reduction of ADCC and CDC. The bulky structure of the nonreducing end of N-linked Fc oligosaccharides is considered to contribute the CDC change, whereas the structural change in the reducing end, i.e. the removal of core fucose, causes ADCC enhancement through improved FcgammaRIIIa binding. In the pharmacokinetic profile, although no significant difference of human neonatal Fc receptor (FcRn)-binding affinity was observed among the three types, the complex type showed longer serum half-lives than the other types irrespective of core fucosylation in mice, which also suggests the contribution of the nonreducing end structure. The present study provides basic information on the effects of core fucose-lacking N-linked Fc oligosaccharides on antibody biological activities.

摘要

已证明,连接到人免疫球蛋白G1(IgG1)抗体恒定区(Fc)的天冬酰胺连接寡糖的结构会影响抗体依赖性细胞毒性(ADCC)和补体依赖性细胞毒性(CDC)的药代动力学及抗体效应功能。然而,N-连接寡糖结构的差异如何影响抗体的生物学活性,尤其是那些缺乏核心岩藻糖的抗体,目前仍不清楚。在此,我们成功地使用同一个生产克隆,生成了具有三种不同N-连接Fc寡糖(即高甘露糖型、杂合型和复合型)且缺乏核心岩藻糖的人IgG1抗体,并比较了它们的活性。在存在或不存在糖苷酶抑制剂(苦马豆素或衣霉素)的情况下,培养α-1,6-岩藻糖基转移酶(FUT8)敲除的中国仓鼠卵巢细胞系,可产生三种类型的抗体,且互不污染。三种非天然存在的非典型寡糖IgG1中,除复合型外,有两种降低了对人淋巴细胞受体IIIa(FcγRIIIa)和补体C1q成分的亲和力,导致ADCC和CDC降低。N-连接Fc寡糖非还原端的庞大结构被认为是导致CDC变化的原因,而还原端的结构变化,即核心岩藻糖的去除,通过改善FcγRIIIa结合导致ADCC增强。在药代动力学方面,尽管三种类型之间未观察到人类新生儿Fc受体(FcRn)结合亲和力的显著差异,但无论小鼠是否存在核心岩藻糖基化,复合型的血清半衰期均比其他类型长,这也表明了非还原端结构的作用。本研究提供了关于缺乏核心岩藻糖的N-连接Fc寡糖对抗体生物学活性影响的基础信息。

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