Directed evolution of potent neutralizing nanobodies against SARS-CoV-2 using CDR-swapping mutagenesis.
Directed evolution of potent neutralizing nanobodies against SARS-CoV-2 using CDR-swapping mutagenesis.
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DOI：
10.1016/j.chembiol.2021.05.019
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发表时间：
2021-09-16
影响因子：
8.6
通讯作者：
Tessier PM
中科院分区：
文献类型：
--
作者：
Zupancic JM;Desai AA;Schardt JS;Pornnoppadol G;Makowski EK;Smith MD;Kennedy AA;Garcia de Mattos Barbosa M;Cascalho M;Lanigan TM;Tai AW;Tessier PM
人们普遍对产生针对SARS-CoV-2和相关病毒的有效中和抗体、纳米抗体和其他亲和性蛋白以应对当前和未来的大流行的简便方法感兴趣。虽然从动物和人类身上分离抗体是行之有效的方法，但这些方法仅限于免疫系统产生的抗体的亲和力、特异性和功能活性。在这里，我们报告了一种令人惊讶的简单的定向进化方法，用于产生具有高亲和力和抗SARS-CoV-2中和活性的纳米体。我们证明了低亲和力铅纳米体之间的互补决定区域交换，我们无意中发现了这一点，但发现这种交换很容易系统地实现，导致成熟的纳米体具有异常大的亲和力增加。重要的是，成熟的纳米体能有效中和SARS-CoV-2假病毒和活病毒，并具有药物样生物物理性质。我们预计，我们的方法将改进体外纳米体发现，并加速产生针对不同冠状病毒的有效中和纳米体。Zupancic等人。报告针对SARS-CoV-2的有效中和纳米抗体。他们展示了一种方法，涉及交换低亲和力克隆的互补决定区域，以产生亲和力和中和活性大幅增加的成熟纳米体。
There is widespread interest in facile methods for generating potent neutralizing antibodies, nanobodies, and other affinity proteins against SARS-CoV-2 and related viruses to address current and future pandemics. While isolating antibodies from animals and humans are proven approaches, these methods are limited to the affinities, specificities, and functional activities of antibodies generated by the immune system. Here we report a surprisingly simple directed evolution method for generating nanobodies with high affinities and neutralization activities against SARS-CoV-2. We demonstrate that complementarity-determining region swapping between low-affinity lead nanobodies, which we discovered unintentionally but find is simple to implement systematically, results in matured nanobodies with unusually large increases in affinity. Importantly, the matured nanobodies potently neutralize both SARS-CoV-2 pseudovirus and live virus, and possess drug-like biophysical properties. We expect that our methods will improve in vitro nanobody discovery and accelerate the generation of potent neutralizing nanobodies against diverse coronaviruses. Zupancic et al. report potent neutralizing nanobodies against SARS-CoV-2. They demonstrate an approach that involves swapping the complementarity-determining regions of low-affinity clones to generate matured nanobodies with large increases in affinity and neutralization activity.
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DOI：
10.1056/nejmoa2029849
发表时间：
2021-01-21
期刊：
The New England journal of medicine
影响因子：
--
作者：
Chen P;Nirula A;Heller B;Gottlieb RL;Boscia J;Morris J;Huhn G;Cardona J;Mocherla B;Stosor V;Shawa I;Adams AC;Van Naarden J;Custer KL;Shen L;Durante M;Oakley G;Schade AE;Sabo J;Patel DR;Klekotka P;Skovronsky DM;BLAZE-1 Investigators
通讯作者：
BLAZE-1 Investigators
影响因子：
64.8
作者：
Barnes CO;Jette CA;Abernathy ME;Dam KA;Esswein SR;Gristick HB;Malyutin AG;Sharaf NG;Huey-Tubman KE;Lee YE;Robbiani DF;Nussenzweig MC;West AP Jr;Bjorkman PJ
通讯作者：
Bjorkman PJ
影响因子：
46.9
作者：
通讯作者：
--
DOI：
10.4049/jimmunol.2000583
发表时间：
2020-08-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Alsoussi WB;Turner JS;Case JB;Zhao H;Schmitz AJ;Zhou JQ;Chen RE;Lei T;Rizk AA;McIntire KM;Winkler ES;Fox JM;Kafai NM;Thackray LB;Hassan AO;Amanat F;Krammer F;Watson CT;Kleinstein SH;Fremont DH;Diamond MS;Ellebedy AH
通讯作者：
Ellebedy AH
影响因子：
16.8
作者：
Huo, Jiangdong;Le Bas, Audrey;Naismith, James H.
通讯作者：
Naismith, James H.