Despite effective countermeasures, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) persists worldwide because of its ability to diversify and evade human immunity. This evasion stems from amino acid substitutions, particularly in the receptor binding domain (RBD) of the spike protein that confers resistance to vaccine-induced antibodies and antibody therapeutics. To constrain viral escape through resistance mutations, we combined antibody variable regions that recognize different RBD sites into multispecific antibodies. Here, we describe multispecific antibodies, including a trivalent trispecific antibody that potently neutralized diverse SARS-CoV-2 variants and prevented virus escape more effectively than single antibodies or mixtures of the parental antibodies. Despite being generated before the appearance of Omicron, this trispecific antibody neutralized all major Omicron variants through BA.4/BA.5 at nanomolar concentrations. Negative stain electron microscopy suggested that synergistic neutralization was achieved by engaging different epitopes in specific orientations that facilitated binding across more than one spike protein. Moreover, a tetravalent trispecific antibody containing the same variable regions as the trivalent trispecific antibody also protected Syrian hamsters against Omicron variants BA.1, BA.2, and BA.5 challenge, each of which uses different amino acid substitutions to mediate escape from therapeutic antibodies. These results demonstrated that multispecific antibodies have the potential to provide broad SARS-CoV-2 coverage, decrease the likelihood of escape, simplify treatment, and provide a strategy for antibody therapies that could help eliminate pandemic spread for this and other pathogens.

中文翻译：

---

针对 SARS-CoV-2 的多特异性抗体可在体外防止免疫逃逸，并在体内提供预防性保护


尽管采取了有效的对策，但严重急性呼吸系统综合症冠状病毒 2 （SARS-CoV-2） 因其具有多样化和逃避人类免疫力的能力而在全球范围内持续存在。这种逃避源于氨基酸替换，特别是在刺突蛋白的受体结合域 （RBD） 中，该结构域赋予对疫苗诱导的抗体和抗体治疗剂的耐药性。为了限制病毒通过耐药突变逃逸，我们将识别不同 RBD 位点的抗体可变区组合成多特异性抗体。在这里，我们描述了多特异性抗体，包括一种三价三特异性抗体，它比单一抗体或亲本抗体的混合物更有效地中和了不同的 SARS-CoV-2 变体并防止病毒逃逸。尽管是在 Omicron 出现之前产生的，但这种三特异性抗体通过纳摩尔浓度的 BA.4/BA.5 中和了所有主要的 Omicron 变体。负染色电子显微镜表明，协同中和是通过在特定方向上结合不同的表位来实现的，这促进了跨多个刺突蛋白的结合。此外，包含与三价三特异性抗体相同可变区的四价三特异性抗体也保护叙利亚仓鼠免受 Omicron 变体 BA.1、BA.2 和 BA.5 攻击，每种变体使用不同的氨基酸取代来介导从治疗性抗体中逃逸。这些结果表明，多特异性抗体有可能提供广泛的 SARS-CoV-2 覆盖，降低逃逸的可能性，简化治疗，并提供一种抗体疗法策略，有助于消除这种病原体和其他病原体的大流行传播。