K E Kilpatrick, D P Danger, E A Hull-Ryde, W Dallas
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引用次数: 14
摘要
在使用Powderject基因枪仅使用5 μ g DNA进行多核苷酸免疫后，在不到4周的时间内生成并分离出了类切换，亲和成熟的小鼠单克隆抗体(MAb)产生与PED/PEA-15反应的细胞系。在开始递送编码目标抗原的DNA后13天进行外周淋巴结细胞的体细胞融合。所提供的数据证明了结合PED/PEA-15的类切换、亲和成熟单克隆抗体的快速生产、鉴定和表征。报道的策略使单克隆抗体的快速开发在酶联免疫吸附测定(ELISA)、Western blotting和免疫沉淀中有用。
High-affinity monoclonal antibodies to PED/PEA-15 generated using 5 microg of DNA.
Class-switched, affinity-matured murine monoclonal antibody (MAb) producing cell lines reactive with PED/PEA-15 were generated and isolated in less than 4 weeks following polynucleotide immunizations using only 5 microg of DNA in conjunction with the Powderject gene gun. Somatic fusions of peripheral lymph node cells were performed 13 days after initiating delivery of DNA encoding the target antigen. The data presented demonstrates the rapid production, identification, and characterization of class-switched, affinity-matured MAbs that bind PED/PEA-15. The reported strategy enabled the rapid development of MAbs that are useful in enzyme-linked immunoadsorbent assay (ELISA), Western blotting, and immunoprecipitations.
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