一种抗CXCL4新单克隆抗体的产生与鉴定
Generation and Characterization of a New Monoclonal Antibody Against CXCL4.
作者信息
Gao Jing, Wu Mingyuan, Gao Jin, Wang Xia, Zhang Yang, Zhu Shunying, Yu Yan, Han Wei
机构信息
1 Laboratory of Regeneromics, School of Pharmacy, School of Agriculture and Biology, Shanghai Jiao Tong University , Shanghai, China .
出版信息
Monoclon Antib Immunodiagn Immunother. 2015 Apr;34(2):110-5. doi: 10.1089/mab.2014.0050.
CXCL4 plays important roles in numerous disease processes, which makes the CXCL4 signaling pathway a potential therapeutic target. In this study, we aimed to develop a neutralizing antibody against both human and mouse CXCL4. Rats were immunized with recombinant human CXCL4 (rhCXCL4). Hybridoma clones were created by fusion of the immunized rat spleen cells with mouse myeloma SP2/0 cells and screened using recombinant mouse CXCL4 (rmCXCL4) and rhCXCL4. The CXCL4 monoclonal antibody (CXCL4 MAb) produced by the 16D6-3 hybridoma clone was sequenced and characterized by Western blot and Biacore assays. It recognized both human and mouse CXCL4 with high affinity and neutralized the effect of rhCXCL4 in vitro. Thus, the antibody may be used in the studies of CXCL4 in murine disease models and as a template in the antibody humanization for clinical developments.
CXCL4在众多疾病过程中发挥着重要作用，这使得CXCL4信号通路成为一个潜在的治疗靶点。在本研究中，我们旨在开发一种针对人和小鼠CXCL4的中和抗体。用重组人CXCL4（rhCXCL4）免疫大鼠。通过将免疫的大鼠脾细胞与小鼠骨髓瘤SP2/0细胞融合来创建杂交瘤克隆，并使用重组小鼠CXCL4（rmCXCL4）和rhCXCL4进行筛选。对16D6-3杂交瘤克隆产生的CXCL4单克隆抗体（CXCL4 MAb）进行测序，并通过蛋白质印迹和生物传感器分析进行表征。它以高亲和力识别人和小鼠CXCL4，并在体外中和rhCXCL4的作用。因此，该抗体可用于小鼠疾病模型中CXCL4的研究，并作为临床开发中抗体人源化的模板。
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