Hayashi Gosuke, Naito Toshinori, Miura Sayaka, Iwamoto Naoya, Usui Yusuke, Bando-Shimizu Mika, Suzuki Sae, Higashi Katsuaki, Nonaka Motohiro, Oishi Shinya, Murakami Hiroshi
Department of Biomolecular Engineering, Graduate School of Engineering, Nagoya University, Nagoya, Japan.
Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto, Japan.
Nat Commun. 2024 Dec 23;15(1):10723. doi: 10.1038/s41467-024-54902-x.
Biologically produced protein drugs are generally susceptible to degradation by proteases and often exhibit immunogenicity. To address this issue, mirror-image peptide/protein binders consisting of D-amino acids have been developed so far through the mirror-image phage display technique. Here, we develop a mirror-image protein binder derived from a monobody, one of the promising protein scaffolds, utilizing two notable technologies: chemical protein synthesis and TRAP display, an improved version of mRNA display. A sequential workflow of initial screening followed by affinity maturation, facilitated by TRAP display, generates an L-monobody with high affinity (K = 1.3 nM) against monocyte chemoattractant protein-1 (MCP-1) D-enantiomer. The chemically synthesized D-monobody demonstrates strong and specific binding to L-MCP-1 and exhibits pharmaceutically favorable properties such as proteolytic resistance, minimal immune response, and a potent inhibitory effect on MCP-1-induced cell migration. This study elevates the value of mirror-image peptide/protein binders as an alternative modality in drug discovery.
生物生产的蛋白质药物通常易受蛋白酶降解，且常表现出免疫原性。为解决这一问题，到目前为止，已通过镜像噬菌体展示技术开发出由D-氨基酸组成的镜像肽/蛋白质结合物。在此，我们利用两种显著技术：化学蛋白质合成和TRAP展示（mRNA展示的改进版本），开发了一种源自单域抗体（一种有前景的蛋白质支架）的镜像蛋白质结合物。由TRAP展示促成的初始筛选随后进行亲和力成熟的连续工作流程，产生了一种对单核细胞趋化蛋白-1（MCP-1）D-对映体具有高亲和力（K = 1.3 nM）的L-单域抗体。化学合成的D-单域抗体表现出与L-MCP-1的强特异性结合，并展现出诸如抗蛋白水解、最小免疫反应以及对MCP-1诱导的细胞迁移有强效抑制作用等药学上有利的特性。这项研究提升了镜像肽/蛋白质结合物作为药物发现中一种替代模式的价值。
