Department of Immunology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Immunotherapy. 2013 Jul;5(7):703-15. doi: 10.2217/imt.13.67.
Immunotherapy with anti-HER2 antibodies has shown promising results in patients with HER2-positive breast cancer. We have recently reported characterization of a mouse monoclonal antibody (mAb) against HER2, which binds to an epitope different from that recognized by trastuzumab and specifically inhibits proliferation of tumor cells overexpressing HER2. In the present study we report chimerization of this antibody.
MATERIALS & METHODS: The immunoglobulin variable region heavy and light chain genes of 1T0 hybridoma cells were amplified and ligated to human γ-1 and κ constant region genes using splice overlap extension PCR. The chimeric antibody was subsequently expressed and characterized by ELISA, western blot and flow cytometry.
The purified chimeric antibody specifically binds to recombinant HER2 and HER2-overexpressing tumor cells and inhibits proliferation of these cells. The binding affinity of the chimeric mAb was comparable with the parental mouse mAb.
This chimeric anti-HER2 mAb is a potentially valuable tool for targeted immunotherapy.
抗 HER2 抗体的免疫疗法已在 HER2 阳性乳腺癌患者中显示出良好的效果。我们最近报道了一种针对 HER2 的鼠单克隆抗体（mAb）的鉴定，该抗体与曲妥珠单抗识别的表位不同，并且特异性抑制过表达 HER2 的肿瘤细胞的增殖。在本研究中，我们报告了该抗体的嵌合化。
使用拼接重叠延伸 PCR，从 1T0 杂交瘤细胞中扩增免疫球蛋白可变区重链和轻链基因，并将其连接到人 γ-1 和 κ 恒定区基因上。随后通过 ELISA、western blot 和流式细胞术对嵌合抗体进行表达和鉴定。
纯化的嵌合抗体特异性结合重组 HER2 和过表达 HER2 的肿瘤细胞，并抑制这些细胞的增殖。嵌合 mAb 的结合亲和力与亲本鼠 mAb 相当。
这种嵌合抗 HER2 mAb 是一种有潜力的靶向免疫治疗工具。
