嵌合抗癌胚抗原单克隆抗体及其Fab片段的研发与特性分析
Development and characterization of chimeric anti-carcinoembryonic antigen monoclonal antibodies and their Fab fragments.
作者信息
Kamigaki T, Ohyanagi H, Yamamoto M, Kaneda T, Goto T, Ohmura T, Yokoyama K, Saitoh Y
机构信息
First Department of Surgery, Kobe University School of Medicine.
出版信息
Jpn J Cancer Res. 1994 Mar;85(3):298-305. doi: 10.1111/j.1349-7006.1994.tb02097.x.
In an attempt to reduce the immunogenicity of two different murine anti-carcinoembryonic antigen (CEA) monoclonal antibodies (MAbs), KM10 and A10, we produced recombinant mouse/human chimeric MAbs and the respective Fab fragments carrying the variable regions of the murine MAbs. Chimeric A10 Fab fragment was expressed in Escherichia coli, and produced in large quantities in a mini-jar fermentation system. In competitive binding assays, chimeric MAbs and their Fab fragments showed identical specificity to human CEA epitopes, as compared to the parental MAbs or Fab fragments. Both chimeric Fab fragments exhibited strong immunohistochemical reactivity with various gastrointestinal carcinomas and no reactivity with CEA-related antigens, such as NCA (nonspecific cross-reacting antigen) or BGPI (biliary glycoprotein I). Furthermore, chimeric KM10 MAb elicited substantially higher antibody-dependent cellular cytotoxicity than the murine MAb. Complement-dependent cytotoxicity in vitro was much weaker with chimeric KM10 MAb. These results indicate that chimeric MAbs or Fab fragments could potentially replace the parental murine antibodies or their Fab fragments in therapy or diagnosis of human gastrointestinal carcinomas.
为降低两种不同的鼠抗癌胚抗原（CEA）单克隆抗体（MAb）KM10和A10的免疫原性，我们制备了重组小鼠/人嵌合单克隆抗体以及携带鼠单克隆抗体可变区的相应Fab片段。嵌合A10 Fab片段在大肠杆菌中表达，并在小型罐发酵系统中大量生产。在竞争性结合试验中，与亲本单克隆抗体或Fab片段相比，嵌合单克隆抗体及其Fab片段对人CEA表位具有相同的特异性。两种嵌合Fab片段对各种胃肠道癌均表现出强烈的免疫组织化学反应，而与CEA相关抗原如NCA（非特异性交叉反应抗原）或BGPI（胆汁糖蛋白I）无反应。此外，嵌合KM10单克隆抗体引发的抗体依赖性细胞毒性比鼠单克隆抗体高得多。嵌合KM10单克隆抗体在体外的补体依赖性细胞毒性要弱得多。这些结果表明，嵌合单克隆抗体或Fab片段在人类胃肠道癌的治疗或诊断中可能会取代亲本鼠抗体或其Fab片段。
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