Abstract：
Resistance to therapeutic antibodies caused by on-target point mutations is a major obstacle in anticancer therapy, creating an \"unmet clinical need.\" To tackle this problem, researchers are developing new generations of antibody drugs that can overcome the resistance mechanisms of existing agents. We have previously reported a structure-guided and phage-assisted evolution (SGAPAE) approach to evolve cetuximab, a therapeutic antibody, to effectively reverse the resistance driven by EGFRS492R or EGFRG465R mutations, without changing the binding epitope or compromising the antibody efficacy. In this protocol, we provide detailed instructions on how to use the SGAPAE approach to evolve cetuximab, which can also be applied to other therapeutic antibodies for reversing on-target point mutation-mediated resistance. The protocol consists of four steps: structure preparation, computational prediction, phage display library construction, and antibody candidate selection.
摘要：
靶向点突变引起的对治疗性抗体的抗性是抗癌治疗的主要障碍,创造了一个“未满足的临床需求”。“为了解决这个问题，研究人员正在开发新一代的抗体药物，可以克服现有药物的耐药机制。我们以前报道了一种结构指导和噬菌体辅助进化（SGAPAE）方法来进化西妥昔单抗，一种治疗性抗体，有效逆转由EGFRS492R或EGFRG465R突变驱动的抗性，而不改变结合表位或损害抗体功效。在这个协议中,我们提供了关于如何使用SGAPAE方法进化西妥昔单抗的详细说明，它也可以应用于其他治疗性抗体，用于逆转靶点突变介导的抗性。该协议包括四个步骤：结构准备，计算预测，噬菌体展示文库的构建,和抗体候选物选择。
