Department of Applied Genetics and Cell Biology, Institute of Plant Biotechnology and Cell Biology, University of Natural Resources and Life Sciences Vienna, Vienna, Austria.
Department of Chemistry, Institute of Biochemistry, University of Natural Resources and Life Sciences Vienna, Vienna, Austria.
Biotechnol J. 2021 Jun;16(6):e2000566. doi: 10.1002/biot.202000566. Epub 2021 Feb 12.
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.
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。高浓度可溶性 ACE2 的给药可被用作诱饵来阻断病毒与细胞 ACE2 受体的相互作用，并且可能被用作治疗或预防 2019 年冠状病毒病的策略。人 ACE2 高度糖基化，其聚糖影响与 SARS-CoV-2 刺突蛋白的结合和病毒感染力。在这里，我们描述了在糖基化工程的烟草中生产重组可溶性 ACE2-片段结晶（Fc）变体。我们的数据表明，产生的二聚体 ACE2-Fc 变体主要与复杂的人类型 N-聚糖糖基化，并且在酶活性、与 SARS-CoV-2 受体结合域的亲和力以及对野生型病毒的中和作用方面具有功能。
