Division of Medical Devices, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo, 158-8501, Japan.
Cytotechnology. 2001 Jul;36(1-3):101-7. doi: 10.1023/A:1014060113186.
We reported previously that adenocarcinoma-reactive human monoclonal antibody AE6F4, which had been generated by in vitro immunization method, recognizes both 14-3-3protein and cytokeratin 8 (CK8). In this study, to analyze the cross-reactivity of AE6F4 antibody, epitopes of AE6F4 antibody on 14-3-3 proteins and CK8 were studied by using synthetic linear peptide scanning technology. To determine the locations of B cell epitope, 48 and 95 of decapeptides covering the entire 14-3-3 proteins and CK8, respectively,were synthesized and binding to AE6F4 antibody was examined by ELISA. The AE6F4 antibody was strongly reactive to peptides containing amino acid sequences TLWTSDTQGD in 14-3-3 proteins and INFLRQLYEE in CK8. These results indicate that AE6F4 antibody can recognize the different peptide sequences in 14-3-3 proteins and CK8.
我们之前曾报道过，通过体外免疫方法产生的腺癌反应性人源单克隆抗体 AE6F4 可识别 14-3-3 蛋白和细胞角蛋白 8（CK8）。在这项研究中，为了分析 AE6F4 抗体的交叉反应性，我们使用合成线性肽扫描技术研究了 AE6F4 抗体在 14-3-3 蛋白和 CK8 上的表位。为了确定 B 细胞表位的位置，合成了覆盖整个 14-3-3 蛋白和 CK8 的分别包含 48 个和 95 个十肽的肽，并通过 ELISA 检查它们与 AE6F4 抗体的结合情况。AE6F4 抗体与 14-3-3 蛋白中的 TLWTSDTQGD 氨基酸序列和 CK8 中的 INFLRQLYEE 氨基酸序列的肽强烈反应。这些结果表明，AE6F4 抗体可以识别 14-3-3 蛋白和 CK8 中的不同肽序列。
