Selection, biophysical and structural analysis of synthetic nanobodies that effectively neutralize SARS-CoV-2.
Selection, biophysical and structural analysis of synthetic nanobodies that effectively neutralize SARS-CoV-2.
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有效中和SARS-CoV-2的合成纳米体的选择、生物物理和结构分析
DOI：
10.1038/s41467-020-19204-y
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发表时间：
2020-11-04
影响因子：
16.6
通讯作者：
Löw C
中科院分区：
文献类型：
--
作者：
Custódio TF;Das H;Sheward DJ;Hanke L;Pazicky S;Pieprzyk J;Sorgenfrei M;Schroer MA;Gruzinov AY;Jeffries CM;Graewert MA;Svergun DI;Dobrev N;Remans K;Seeger MA;McInerney GM;Murrell B;Hällberg BM;Löw C
冠状病毒SARS-CoV-2是持续的COVID-19大流行的原因。治疗性中和抗体是应对COVID-19的关键中短期方法。然而，传统的抗体生产受到开发时间长和生产成本高的阻碍。在这里，我们报告了从合成库中快速分离和表征纳米抗体，称为合成抗体（Sb），其靶向SARS-CoV-2刺突蛋白的受体结合结构域（RBD）。鉴定了几种具有低纳摩尔亲和力和有效中和活性的结合剂，其中Sb 23显示出高亲和力并中和假病毒，IC 50为0.6 µg/ml。与Sb 23结合的刺突的冷冻电镜结构表明Sb 23在ACE 2结合位点竞争性结合。此外，cryo-EM重建揭示了一个不寻常的构象的尖峰，其中两个RBD是在'向上' ACE 2结合构象。该组合方法代表了一种替代的快速工作流程，以选择对新出现的病毒具有中和活性的结合剂。在这里，作者从一个合成文库中分离出几种纳米抗体，它们结合SARS-CoV-2刺突蛋白（S）的受体结合域（RBD）并中和S假型病毒。具有一个纳米抗体的刺突的Cryo-EM结构和进一步的生物物理分析显示与ACE 2结合的竞争。
The coronavirus SARS-CoV-2 is the cause of the ongoing COVID-19 pandemic. Therapeutic neutralizing antibodies constitute a key short-to-medium term approach to tackle COVID-19. However, traditional antibody production is hampered by long development times and costly production. Here, we report the rapid isolation and characterization of nanobodies from a synthetic library, known as sybodies (Sb), that target the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein. Several binders with low nanomolar affinities and efficient neutralization activity were identified of which Sb23 displayed high affinity and neutralized pseudovirus with an IC50 of 0.6 µg/ml. A cryo-EM structure of the spike bound to Sb23 showed that Sb23 binds competitively in the ACE2 binding site. Furthermore, the cryo-EM reconstruction revealed an unusual conformation of the spike where two RBDs are in the ‘up’ ACE2-binding conformation. The combined approach represents an alternative, fast workflow to select binders with neutralizing activity against newly emerging viruses. Here, the authors isolate several nanobodies from a synthetic library that bind the receptor-binding domain (RBD) of SARS-CoV-2 spike protein (S) and neutralize S pseudotyped viruses. Cryo-EM structure of Spike with one nanobody and further biophysical analysis shows competition with ACE2 binding.
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影响因子：
14.8
作者：
Kelley LA;Mezulis S;Yates CM;Wass MN;Sternberg MJ
通讯作者：
Sternberg MJ
影响因子：
48
作者：
Punjani, Ali;Rubinstein, John L.;Brubaker, Marcus A.
通讯作者：
Brubaker, Marcus A.
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH
影响因子：
5.2
作者：
Ortega C;Prieto D;Abreu C;Oppezzo P;Correa A
通讯作者：
Correa A
影响因子：
6.1
作者：
Blanchet CE;Spilotros A;Schwemmer F;Graewert MA;Kikhney A;Jeffries CM;Franke D;Mark D;Zengerle R;Cipriani F;Fiedler S;Roessle M;Svergun DI
通讯作者：
Svergun DI