Department of Chemical Engineering, University of Michigan, North Campus Research Complex, 2800 Plymouth Road, Ann Arbor, MI 48109, USA; Biointerfaces Institute, University of Michigan, Ann Arbor, MI 48109, USA.
Department of Chemical Engineering, University of Michigan, North Campus Research Complex, 2800 Plymouth Road, Ann Arbor, MI 48109, USA; Department of Pharmaceutical Sciences, University of Michigan, Ann Arbor, MI 48109, USA; Biointerfaces Institute, University of Michigan, Ann Arbor, MI 48109, USA.
Cell Chem Biol. 2021 Sep 16;28(9):1379-1388.e7. doi: 10.1016/j.chembiol.2021.05.019. Epub 2021 Jun 24.
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.
人们普遍关注能够简便地生成针对 SARS-CoV-2 及相关病毒的强效中和抗体、纳米抗体和其他亲和蛋白的方法，以应对当前和未来的大流行。虽然从动物和人类中分离抗体是已被证实的方法，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，用于生成针对 SARS-CoV-2 具有高亲和力和中和活性的纳米抗体。我们证明，在我们无意中发现但发现可以系统地简单实施的低亲和力先导纳米抗体之间进行互补决定区交换，会导致亲和力大幅提高的成熟纳米抗体。重要的是，成熟的纳米抗体能够有效中和 SARS-CoV-2 假病毒和活病毒，并具有类药的物理化学特性。我们预计我们的方法将改进体外纳米抗体的发现，并加速生成针对多种冠状病毒的强效中和纳米抗体。
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