Liu Chao, Xu Hongrong, Yuan Fei, Chen Weili, Chen Hanjing, Sheng Lei, Wu Yujia, Li Xuening
Department of Clinical Pharmacology, Zhongshan Hospital, Fudan University, 200032, Shanghai, China.
Department of Clinical Pharmacology, Zhongshan Hospital, Fudan University, 200032, Shanghai, China.
Biochem Biophys Res Commun. 2022 May 28;606:55-60. doi: 10.1016/j.bbrc.2022.03.066. Epub 2022 Mar 16.
Proprotein convertase subtilisin/kexin type 9 (PCSK9), a negative regulator of LDLR, has emerged as an important target for the treatment of hypercholesterolemic cardiovascular disease, and monoclonal antibodies alirocumab and evolocumab against it have been widely used in clinical practice. The vaccine research of PCSK9 is considered a promising option for the long-term treatment and prevention of cardiovascular disease, but progress has been slow. The selection of safe and effective epitopes is one of the key steps in vaccine development. In this study, we designed a phage display library of cascaded peptides for affinity screening with two antibody drugs, and found that the two peptides PC3 and PS6, which are adjacent to each other in protein spatial structure, both have superior binding activity to the screening antibodies. We performed in vitro recombination design on the dominant sequences, and obtained recombinant sequences that can respond to the dominant conformational epitope of PCSK9, which provides a meaningful reference for epitope selection in subsequent PCSK9 vaccine development.
前蛋白转化酶枯草溶菌素/kexin 9型（PCSK9）是低密度脂蛋白受体（LDLR）的负调节因子，已成为治疗高胆固醇血症心血管疾病的重要靶点，针对它的单克隆抗体阿利西尤单抗和依洛尤单抗已在临床实践中广泛应用。PCSK9疫苗研究被认为是心血管疾病长期治疗和预防的一个有前景的选择，但进展缓慢。选择安全有效的表位是疫苗开发的关键步骤之一。在本研究中，我们设计了一个级联肽噬菌体展示文库，用于与两种抗体药物进行亲和筛选，发现两种在蛋白质空间结构上相邻的肽PC3和PS6对筛选抗体均具有优异的结合活性。我们对优势序列进行了体外重组设计，获得了能够响应PCSK9优势构象表位的重组序列，这为后续PCSK9疫苗开发中的表位选择提供了有意义的参考。
