Many reports have indicated that the insulin receptor (IR) causes tumorigenesis and the development of breast cancer. It has been considered a potential target for treating IR-related tumors. Traditionally, there are two categories of insulin receptor (IR) antagonists, they are small molecule antagonists and anti-IR antibodies. Here, we describe a new method (anti-idiotypic antibody strategy) for the development of IR antagonist. Hybridoma technology was employed to design and identify a series of anti-idiotypic antibodies against insulin. After repeated screening and identification, an anti-idiotypic antibody against IR (AK98) was obtained. Analysis through competitive ELISA and competitive receptor binding indicated that AK98 mimicked the receptor binding epitope of insulin. The interaction between AK98 and IR was determined using indirect immunofluorescence, immunoelectron microscopy, and Immunoprecipitation-Western (IP-WB). Further research using a tumor cell model revealed that AK98 inhibited insulin-IR binding and IR-mediated intracellular signaling pathways. Conclusively, the main purpose of this paper is that we proposed a new method (anti-idiotypic antibody strategy) to develop the insulin receptor (IR) antagonist (AK98), and a series of experiments showed that the anti-idiotypic antibody (AK98) exhibited good antagonistic activity against IR. This work suggests that the anti-idiotypic antibody may be a potential strategy to develop IR antagonists that can be used in treating breast cancer.

许多报告表明，胰岛素受体（IR）会导致肿瘤的发生和乳腺癌的发展。已经认为它是治疗IR相关肿瘤的潜在靶标。传统上，胰岛素受体（IR）拮抗剂有两类，它们是小分子拮抗剂和抗IR抗体。在这里，我们描述了开发IR拮抗剂的新方法（抗独特型抗体策略）。杂交瘤技术被用于设计和鉴定一系列针对胰岛素的抗独特型抗体。经过反复筛选和鉴定后，获得了针对IR的抗独特型抗体（AK98）。通过竞争性ELISA和竞争性受体结合的分析表明，AK98模仿胰岛素的受体结合表位。使用间接免疫荧光，免疫电子显微镜和免疫沉淀-Western（IP-WB）测定AK98和IR之间的相互作用。使用肿瘤细胞模型的进一步研究表明，AK98抑制胰岛素-IR结合和IR介导的细胞内信号通路。总之，本文的主要目的是我们提出了一种开发胰岛素受体（IR）拮抗剂（AK98）的新方法（抗独特型抗体策略），并且一系列实验表明该抗独特型抗体（AK98）对IR表现出良好的拮抗活性。这项工作表明，抗独特型抗体可能是开发可用于治疗乳腺癌的IR拮抗剂的潜在策略。使用肿瘤细胞模型的进一步研究表明，AK98抑制胰岛素-IR结合和IR介导的细胞内信号通路。总之，本文的主要目的是我们提出了一种开发胰岛素受体（IR）拮抗剂（AK98）的新方法（抗独特型抗体策略），并且一系列实验表明该抗独特型抗体（AK98）对IR表现出良好的拮抗活性。这项工作表明，抗独特型抗体可能是开发可用于治疗乳腺癌的IR拮抗剂的潜在策略。使用肿瘤细胞模型的进一步研究表明，AK98抑制胰岛素-IR结合和IR介导的细胞内信号通路。总之，本文的主要目的是我们提出了一种开发胰岛素受体（IR）拮抗剂（AK98）的新方法（抗独特型抗体策略），并且一系列实验表明该抗独特型抗体（AK98）对IR表现出良好的拮抗活性。这项工作表明，抗独特型抗体可能是开发可用于治疗乳腺癌的IR拮抗剂的潜在策略。一系列实验表明抗独特型抗体（AK98）对IR具有良好的拮抗活性。这项工作表明，抗独特型抗体可能是开发可用于治疗乳腺癌的IR拮抗剂的潜在策略。一系列实验表明抗独特型抗体（AK98）对IR具有良好的拮抗活性。这项工作表明，抗独特型抗体可能是开发可用于治疗乳腺癌的IR拮抗剂的潜在策略。