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

被引：27

作者：

Castilho, Alexandra ^{[1
]}

Schwestka, Jennifer ^{[1
]}

Kienzl, Nikolaus F. ^{[1
]}

Vavra, Ulrike ^{[1
]}

Gruenwald-Gruber, Clemens ^{[2
]}

Izadi, Shiva ^{[1
,3
]}

Hiremath, Chaitra ^{[1
]}

Niederhoefer, Janine ^{[4
]}

Laurent, Elisabeth ^{[5
,6
]}

Monteil, Vanessa ^{[7
,8
]}

Mirazimi, Ali ^{[7
,8
]}

Wirnsberger, Gerald ^{[4
]}

Stadlmann, Johannes ^{[2
]}

Stoeger, Eva ^{[1
]}

Mach, Lukas ^{[1
]}

Strasser, Richard ^{[1
]}

机构：

[1] Univ Nat Resources & Life Sci Vienna, Inst Plant Biotechnol & Cell Biol, Dept Appl Genet & Cell Biol, Vienna, Austria

[2] Univ Nat Resources & Life Sci Vienna, Dept Chem, Inst Biochem, Vienna, Austria

[3] Tarbiat Modares Univ, Fac Agr, Dept Biotechnol, Tehran, Iran

[4] Apeiron Biol AG, Campus Vienna Bioctr, Vienna, Austria

[5] Univ Nat Resources & Life Sci Vienna, Dept Biotechnol, Vienna, Austria

[6] Univ Nat Resources & Life Sci Vienna, Core Facil Biomol & Cellular Anal, Vienna, Austria

[7] Karolinska Inst, Dept Lab Med, Unit Clin Microbiol, Stockholm, Sweden

[8] Karolinska Univ Hosp, Stockholm, Sweden

来源：

BIOTECHNOLOGY JOURNAL | 2021年 / 16卷 / 06期

基金：

奥地利科学基金会;

关键词：

COVID-19; glycosylation; posttranslational modification; recombinant protein expression; SARS-CoV-2; GLYCOSYLATION; ANTIBODIES; EXPRESSION;

D O I：

10.1002/biot.202000566

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

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页数：6

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共 17 条

[1] The role of IgG Fc receptors in antibody-dependent enhancement [J].

Bournazos, Stylianos ;

Gupta, Aaron ;

Ravetch, Jeffrey, V .

NATURE REVIEWS IMMUNOLOGY, 2020, 20 (10) :633-643

[2] Generation of Biologically Active Multi-Sialylated Recombinant Human EPOFc in Plants [J].

Castilho, Alexandra ;

Neumann, Laura ;

Gattinger, Pia ;

Strasser, Richard ;

Vorauer-Uhl, Karola ;

Sterovsky, Thomas ;

Altmann, Friedrich ;

Steinkellner, Herta .

PLOS ONE, 2013, 8 (01)

[3] Engineering human ACE2 to optimize binding to the spike protein of SARS coronavirus 2 [J].

Chan, Kui K. ;

Dorosky, Danielle ;

Sharma, Preeti ;

Abbasi, Shawn A. ;

Dye, John M. ;

Kranz, David M. ;

Herbert, Andrew S. ;

Procko, Erik .

SCIENCE, 2020, 369 (6508) :1261-+

[4] Exploring Site-Specific N-Glycosylation of HEK293 and Plant-Produced Human IgA Isotypes [J].

Goeritzer, Kathrin ;

Maresch, Daniel ;

Altmann, Friedrich ;

Obinger, Christian ;

Strasser, Richard .

JOURNAL OF PROTEOME RESEARCH, 2017, 16 (07) :2560-2570

[5] Pharmacokinetics and Pharmacodynamics of Recombinant Human Angiotensin-Converting Enzyme 2 in Healthy Human Subjects [J].

Haschke, Manuel ;

Schuster, Manfred ;

Poglitsch, Marko ;

Loibner, Hans ;

Salzberg, Marc ;

Bruggisser, Marcel ;

Penninger, Joseph ;

Kraehenbuehl, Stephan .

CLINICAL PHARMACOKINETICS, 2013, 52 (09) :783-792

[6] SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor [J].

Hoffmann, Markus ;

Kleine-Weber, Hannah ;

Schroeder, Simon ;

Krueger, Nadine ;

Herrler, Tanja ;

Erichsen, Sandra ;

Schiergens, Tobias S. ;

Herrler, Georg ;

Wu, Nai-Huei ;

Nitsche, Andreas ;

Mueller, Marcel A. ;

Drosten, Christian ;

Poehlmann, Stefan .

CELL, 2020, 181 (02) :271-+

[7] Neutralization of SARS-CoV-2 spike pseudotyped virus by recombinant ACE2-Ig [J].

Lei, Changhai ;

Qian, Kewen ;

Li, Tian ;

Zhang, Sheng ;

Fu, Wenyan ;

Ding, Min ;

Hu, Shi .

NATURE COMMUNICATIONS, 2020, 11 (01)

[8] Novel ACE2-Fc chimeric fusion provides long-lasting hypertension control and organ protection in mouse models of systemic renin angiotensin system activation [J].

Liu, Pan ;

Wysocki, Jan ;

Souma, Tomokazu ;

Ye, Minghao ;

Ramirez, Veronica ;

Zhou, Bisheng ;

Wilsbacher, Lisa D. ;

Quaggin, Susan E. ;

Batlle, Daniel ;

Jin, Jing .

KIDNEY INTERNATIONAL, 2018, 94 (01) :114-125

[9] Inhibition of SARS-CoV-2 Infections in Engineered Human Tissues Using Clinical-Grade Soluble Human ACE2 [J].

Monteil, Vanessa ;

Kwon, Hyesoo ;

Prado, Patricia ;

Hagelkruys, Astrid ;

Wimmer, Reiner A. ;

Stahl, Martin ;

Leopoldi, Alexandra ;

Garreta, Elena ;

del Pozo, Carmen Hurtado ;

Prosper, Felipe ;

Romero, Juan Pablo ;

Wirnsberger, Gerald ;

Zhang, Haibo ;

Slutsky, Arthur S. ;

Conder, Ryan ;

Montserrat, Nuria ;

Mirazimi, Ali ;

Penninger, Josef M. .

CELL, 2020, 181 (04) :905-+

[10] The human anti-HIV antibodies 2F5, 2G12, and PG9 differ in their susceptibility to proteolytic degradation: Down- regulation of endogenous serine and cysteine proteinase activities could improve antibody production in plant- based expression platforms [J].

Niemer, Melanie ;

Mehofer, Ulrich ;

Acosta, Juan Antonio Torres ;

Verdianz, Maria ;

Henkel, Theresa ;

Loos, Andreas ;

Strasser, Richard ;

Maresch, Daniel ;

Rademacher, Thomas ;

Steinkellner, Herta ;

Mach, Lukas .

BIOTECHNOLOGY JOURNAL, 2014, 9 (04) :493-500