受体酪氨酸激酶样孤儿受体 1 (ROR1) 的过度表达有助于癌细胞增殖、存活和迁移，在肿瘤发展中发挥关键作用。 ROR1已被提议作为癌症治疗的潜在治疗靶点。本研究旨在开发新型人源化ROR1单克隆抗体并研究其抗肿瘤作用。采用免疫组化和流式细胞术分析肿瘤组织和细胞系中ROR1的表达。通过互补决定区（CDR）移植技术将小鼠杂交瘤的抗体人源化。采用表面等离子共振光谱、ELISA 测定和流式细胞术来表征人源化抗体。通过体外细胞测定和体内小鼠实验来综合评价这些抗体的抗肿瘤活性。ROR1在肺腺癌、肝癌和乳腺癌中表现出显着较高的表达，短发夹RNA靶向ROR1可显着抑制增殖和迁移癌细胞。成功开发了两种人源化ROR1单克隆抗体，命名为h1B8和h6D4，对ROR1蛋白具有高特异性和亲和力。此外，这两种抗体在肺癌异种移植小鼠模型、c-Myc/Alb-cre肝癌转基因小鼠模型和MMTV-PyMT乳腺癌小鼠模型中均能有效抑制肿瘤生长。针对ROR1、h1B8和h6D4的两种人源化单克隆抗体成功开发并在体内表现出显着的抗肿瘤活性。© 2024。作者。

Overexpression of receptor tyrosine kinase-like orphan receptor 1 (ROR1) contributes to cancer cell proliferation, survival and migration, playing crucial roles in tumor development. ROR1 has been proposed as a potential therapeutic target for cancer treatment. This study aimed to develop novel humanized ROR1 monoclonal antibodies and investigate their anti-tumor effects.ROR1 expression in tumor tissues and cell lines was analyzed by immunohistochemistry and flow cytometry. Antibodies from mouse hybridomas were humanized by the complementarity-determining region (CDR) grafting technique. Surface plasmon resonance spectroscopy, ELISA assay and flow cytometry were employed to characterize humanized antibodies. In vitro cellular assay and in vivo mouse experiment were conducted to comprehensively evaluate anti-tumor activity of these antibodies.ROR1 exhibited dramatically higher expression in lung adenocarcinoma, liver cancer and breast cancer, and targeting ROR1 by short-hairpin RNAs significantly inhibited proliferation and migration of cancer cells. Two humanized ROR1 monoclonal antibodies were successfully developed, named h1B8 and h6D4, with high specificity and affinity to ROR1 protein. Moreover, these two antibodies effectively suppressed tumor growth in the lung cancer xenograft mouse model, c-Myc/Alb-cre liver cancer transgenic mouse model and MMTV-PyMT breast cancer mouse model.Two humanized monoclonal antibodies targeting ROR1, h1B8 and h6D4, were successfully developed and exhibited remarkable anti-tumor activity in vivo.© 2024. The Author(s).