Generation of enzymatically competent SARS-CoV-2 decoy receptor ACE2-Fc in glycoengineered Nicotiana benthamiana,Biotechnology Journal

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Generation of enzymatically competent SARS-CoV-2 decoy receptor ACE2-Fc in glycoengineered Nicotiana benthamiana
Biotechnology Journal ( IF 3.2 ) Pub Date : 2021-01-22 , DOI: 10.1002/biot.202000566
Alexandra Castilho ₁ , Jennifer Schwestka ₁ , Nikolaus F Kienzl ₁ , Ulrike Vavra ₁ , Clemens Grünwald-Gruber ₂ , Shiva Izadi _{1,

3} , Chaitra Hiremath ₁ , Janine Niederhöfer ₄ , Elisabeth Laurent ₅ , Vanessa Monteil ₆ , Ali Mirazimi ₆ , Gerald Wirnsberger ₄ , Johannes Stadlmann ₂ , Eva Stöger ₁ , Lukas Mach ₁ , Richard Strasser ₁

Affiliation

Human angiotensin-converting enzyme 2 (ACE2) is the primary host cell receptor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) binding and cell entry. Administration of high concentrations of soluble ACE2 can be utilized as a decoy to block the interaction of the virus with cellular ACE2 receptors and potentially be used as a strategy for treatment or prevention of coronavirus disease 2019. Human ACE2 is heavily glycosylated and its glycans impact on binding to the SARS-CoV-2 spike protein and virus infectivity. Here, we describe the production of a recombinant soluble ACE2-fragment crystallizable (Fc) variant in glycoengineered Nicotiana benthamiana. Our data reveal that the produced dimeric ACE2-Fc variant is glycosylated with mainly complex human-type N-glycans and functional with regard to enzyme activity, affinity to the SARS-CoV-2 receptor-binding domain, and wild-type virus neutralization.

中文翻译：

在糖工程烟草中产生具有酶活性的 SARS-CoV-2 诱饵受体 ACE2-Fc

人血管紧张素转换酶 2 (ACE2) 是严重急性呼吸综合征冠状病毒 2 (SARS-CoV-2) 结合和进入细胞的主要宿主细胞受体。施用高浓度的可溶性 ACE2 可用作诱饵，阻断病毒与细胞 ACE2 受体的相互作用，并有可能用作治疗或预防 2019 年冠状病毒病的策略。人类 ACE2 高度糖基化，其聚糖对与 SARS-CoV-2 刺突蛋白和病毒感染性的结合。在这里，我们描述了在糖工程烟草中生产重组可溶性 ACE2 片段可结晶 (Fc) 变体。我们的数据显示，产生的二聚体 ACE2-Fc 变体主要与复杂的人型N-聚糖进行糖基化，并在酶活性、对 SARS-CoV-2 受体结合结构域的亲和力以及野生型病毒中和方面具有功能。

更新日期：2021-01-22

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