Preparation, Purification, and Identification of a Monoclonal Antibody Against NRP2 b1b2 Domain
Preparation, Purification, and Identification of a Monoclonal Antibody Against NRP2 b1b2 Domain
复制标题
抗 NRP2 b1b2 结构域单克隆抗体的制备、纯化和鉴定。
DOI：
10.1089/mab.2015.0025
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发表时间：
2015-10-01
影响因子：
--
通讯作者：
Yan, Jiang-Hua
中科院分区：
文献类型：
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作者：
Yang, Yun;Chen, Na;Yan, Jiang-Hua
神经纤毛蛋白2（Neuropilin2，NRP2）是一种跨膜的非酪氨酸激酶糖蛋白，在神经元的形成中起重要作用，它首先被鉴定为3类脑信号蛋白和血管内皮生长因子（VEGF）家族的高亲和力激酶缺陷型受体。此外，NRP2表达通常在癌组织中上调，并与不良预后相关。本文报道了利用杂交瘤技术制备NRP2b1b2结构域单克隆抗体（NRP2MAb）。用间接ELISA法测得NRP2单克隆抗体对NRP2b1b2的效价为5.12 × 10（5）。Western blotting、流式细胞术和免疫荧光分析表明NRP2单克隆抗体可在LoVo和SW 480细胞中与全长NRP2联合收割机结合。除了有助于进一步了解NRP2相关的病理机制和细胞信号通路外，NRP2单克隆抗体未来可能作为癌症的治疗剂。
First identified as a high-affinity kinase-deficient receptor for class-3 semaphorins and vascular endothelial growth factor (VEGF) families, Neuropilin2 (NRP2) is a transmembrane non-tyrosine-kinase glycoprotein that has a vital function in neuronal patterning. Furthermore, NRP2 expression is often upregulated in cancer tissues and correlated with poor prognosis. In the present study, we report the establishment of a monoclonal antibody specific for NRP2b1b2 domain (NRP2 MAb) through hybridoma method. NRP2 MAb is measured to have a titer of 5.12 × 10(5) against NRP2b1b2 in indirect ELISA. Western blotting, flow cytometry, and immunofluorescence analysis indicate that NRP2 MAb can combine full-length NRP2 in LoVo and SW480 cells. Besides helping further understand NRP2-related pathological mechanisms and cell-signaling pathways, NRP2 MAb may act as a therapeutic agent for cancer in the future.