Modulation of therapeutic antibody effector functions by glycosylation engineering:: Influence of Golgi enzyme localization domain and co-expression of heterologous β1,4-N-acetylglucosaminyltransferase III and Golgi α-mannosidase II
Modulation of therapeutic antibody effector functions by glycosylation engineering:: Influence of Golgi enzyme localization domain and co-expression of heterologous β1,4-N-acetylglucosaminyltransferase III and Golgi α-mannosidase II
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DOI：
10.1002/bit.20777
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发表时间：
2006-04-05
影响因子：
3.8
通讯作者：
Umaña, P
中科院分区：
文献类型：
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作者：
Ferrara, C;Brünker, P;Umaña, P
IgG抗体引起的效应子功能强烈依赖于与蛋白质Fc区连接的碳水化合物部分。因此，已经开发了几种方法来合理地操纵这些聚糖并改善抗体的生物学功能。在生产细胞系中过表达重组β 1，4-N-乙酰基-葡糖胺基转移酶III（GnT-III）可产生富含二分寡糖的抗体。此外，GnT-III过表达导致非岩藻糖基化和杂合寡糖增加。此类抗体糖变体具有增加的抗体依赖性细胞毒性（ADCC）。为了探索除了GnT-III过表达之外的另一个变量，我们将GnT-III的定位结构域与其他高尔基体驻留酶的定位结构域进行了交换。我们的结果表明，嵌合GnT-III可以更有效地与内源性核心α 1，6-岩藻糖基转移酶（α 1，6-FucT）和高尔基体α-甘露糖苷酶II（ManII）竞争，导致更高比例的二等分非岩藻糖基化杂合聚糖（“Glyco-1”抗体）。GnT-III和ManII的共表达导致与Glyco-1相似的非岩藻糖基化程度，但大多数与该抗体连接的寡糖（“Glyco-2”）是复合物类型。与未修饰的抗体相比，这些糖变体的ADCC活性显著增加，而与Glyco-2或未修饰的抗体相比，Glyco-1（富含杂交体）的补体依赖性细胞毒性（CDC）降低。我们表明，除了GnT-III过表达，工程GnT-III定位是一个通用的工具，以调节其治疗应用相关的抗体的生物活性。(c)2006 Wiley Periodicals，Inc.
The effector functions elicited by 1gG antibodies strongly depend on the carbohydrate moiety linked to the Fc region of the protein. Therefore several approaches have been developed to rationally manipulate these glycans and improve the biological functions of the antibody. Overexpression of recombinant beta 1,4-N-acetyl-glucosaminyltransferase III (GnT-III) in production cell lines leads to antibodies enriched in bisected oligosaccharides. Moreover, GnT-III overexpression leads to increases in non-fucosylated and hybrid oligosaccharides. Such antibody glycovariants have increased antibody-dependent cellular cytotoxicity (ADCC). To explore a further variable besides overexpression of GnT-III, we exchanged the localization domain of GnT-III with that of other Golgi-resident enzymes. Our results indicate that chimeric GnT-III can compete even more efficiently against the endogenous core alpha 1,6-fucosyltransferase (alpha 1,6-FucT) and Golgi alpha-mannosidase II (ManII) leading to higher proportions of bisected non-fucosylated hybrid glycans ("Glyco-1" antibody). The co-expression of GnT-III and ManII led to a similar degree of non-fucosylation as that obtained for Glyco-1, but the majority of the oligosaccharides linked to this antibody ("Glyco-2") are of the complex type. These glycovariants feature strongly increased ADCC activity compared to the unmodified antibody, while Glyco-1 (hybrid-rich) features reduced complement-dependent cytotoxicity (CDC) compared to Glyco-2 or unmodified antibody. We show that apart from GnT-III overexpression, engineering of GnT-III localization is a versatile tool to modulate the biological activities of antibodies relevant for their therapeutic application. (c) 2006 Wiley Periodicals, Inc.