Department of Surgery, University of Vermont College of Medicine, Burlington, VT 05405, USA.
Cancer Immunol Immunother. 2013 Aug;62(8):1397-410. doi: 10.1007/s00262-013-1443-5. Epub 2013 Jun 5.
Phage display is a powerful method for target discovery and selection of ligands for cancer treatment and diagnosis. Our goal was to select tumor-binding antibodies in cancer patients. Eligibility criteria included absence of preexisting anti-phage-antibodies and a Stage IV cancer status. All patients were intravenously administered 1 × 10(11) TUs/kg of an scFv library 1 to 4 h before surgical resection of their tumors. No significant adverse events related to the phage library infusion were observed. Phage were successfully recovered from all tumors. Individual clones from each patient were assessed for binding to the tumor from which clones were recovered. Multiple tumor-binding phage-antibodies were identified. Soluble scFv antibodies were produced from the phage clones showing higher tumor binding. The tumor-homing phage-antibodies and derived soluble scFvs were found to bind varying numbers (0-5) of 8 tested normal human tissues (breast, cervix, colon, kidney, liver, spleen, skin, and uterus). The clones that showed high tumor-specificity were found to bind corresponding tumors from other patients also. Clone enrichment was observed based on tumor binding and DNA sequence data. Clone sequences of multiple variable regions showed significant matches to certain cancer-related antibodies. One of the clones (07-2,355) that was found to share a 12-amino-acid-long motif with a reported IL-17A antibody was further studied for competitive binding for possible antigen target identification. We conclude that these outcomes support the safety and utility of phage display library panning in cancer patients for ligand selection and target discovery for cancer treatment and diagnosis.
噬菌体展示是一种强大的方法，可用于发现靶标并选择用于癌症治疗和诊断的配体。我们的目标是在癌症患者中选择与肿瘤结合的抗体。入选标准包括无预先存在的抗噬菌体抗体和 IV 期癌症状态。所有患者在手术切除肿瘤前 1 至 4 小时静脉内给予 1×10(11)TU/kg 的 scFv 文库 1。未观察到与噬菌体文库输注相关的严重不良事件。从所有肿瘤中成功回收了噬菌体。从每位患者中评估了单个克隆与从其回收克隆的肿瘤的结合情况。鉴定了多种与肿瘤结合的噬菌体抗体。从显示更高肿瘤结合的噬菌体克隆中产生了可溶性 scFv 抗体。发现肿瘤归巢的噬菌体抗体和衍生的可溶性 scFv 可结合 8 种测试的正常人体组织（乳腺、宫颈、结肠、肾脏、肝脏、脾脏、皮肤和子宫）中的不同数量（0-5）。发现显示高肿瘤特异性的克隆也与其他患者的相应肿瘤结合。基于肿瘤结合和 DNA 序列数据观察到克隆富集。多个可变区的克隆序列与某些与癌症相关的抗体有明显的匹配。发现一个与报道的 IL-17A 抗体共享 12 个氨基酸长的基序的克隆（07-2,355），进一步研究了其竞争结合的可能性，以确定可能的抗原靶标。我们得出结论，这些结果支持在癌症患者中使用噬菌体展示文库淘选进行配体选择和靶标发现用于癌症治疗和诊断的安全性和实用性。
