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首页> 外文期刊>New biotechnology >Generation of HER2 monoclonal antibodies using epitopes of a rabbit polyclonal antibody
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Generation of HER2 monoclonal antibodies using epitopes of a rabbit polyclonal antibody

机译：使用兔多克隆抗体的表位生成HER2单克隆抗体

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One of the issues in using polyclonal antibodies is the limited amount of reagent available from an immunisation, leading to batch-to-batch variation and difficulties in obtaining the same antibody performance when the same antigen is re-immunised into several separate animals. This led to the development of hybridoma technology allowing, at least theoretically, for an unlimited production of a specific binder. Nevertheless, polyclonal antibodies are widely used in research and diagnostics and there exists a need for robust methods to convert a polyclonal antibody with good binding performance into a renewable monoclonal with identical or similar binding specificity. Here we have used precise information regarding the functional recognition sequence (epitope) of a rabbit polyclonal antibody with attractive binding characteristics as the basis for generation of a renewable mouse monoclonal antibody. First, the original protein fragment antigen was used for immunisation and generation of mouse hybridoma, without obtaining binders to the same epitope region. Instead a peptide designed using the functional epitope and structural information was synthesised and used for hybridoma production. Several of the monoclonal antibodies generated were found to have similar binding characteristics to those of the original polyclonal antibody. These monoclonal antibodies detected native HER2 on cell lines and were also able to stain HER2 in immunohistochemistry using xenografted mice, as well as human normal and cancer tissues.
机译：使用多克隆抗体的问题之一是可从免疫中获得的试剂数量有限，导致同一批抗原重新免疫到数只单独的动物中时，批次间变异以及难以获得相同的抗体性能。这导致了杂交瘤技术的发展，至少在理论上允许无限量生产特定的结合剂。然而，多克隆抗体被广泛用于研究和诊断中，并且需要鲁棒的方法将具有良好结合性能的多克隆抗体转化为具有相同或相似结合特异性的可再生单克隆抗体。在这里，我们已使用具有吸引力的结合特性的兔多克隆抗体的功能识别序列（表位）的精确信息，作为生成可再生小鼠单克隆抗体的基础。首先，原始蛋白质片段抗原用于免疫和小鼠杂交瘤的产生，而没有获得相同表位区域的结合物。相反，合成了使用功能性表位和结构信息设计的肽，并将其用于杂交瘤生产。发现产生的几种单克隆抗体具有与原始多克隆抗体相似的结合特性。这些单克隆抗体在细胞系中检测到天然HER2，并且还能够使用异种移植小鼠以及人类正常组织和癌症组织在免疫组织化学中对HER2进行染色。

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