Okamoto S, Yoshikawa K, Obata Y, Shibuya M, Aoki S, Yoshida J, Takahashi T
Laboratory of Immunology, Aichi Cancer Center Research Institute, Nagoya, Japan.
Br J Cancer. 1996 Jun;73(11):1366-72. doi: 10.1038/bjc.1996.260.
A mouse monoclonal antibody (IgG2b), 3C10, was produced against the truncated epidermal growth factor receptor (EGFR), encoded by the (type III) in-frame deletion mutation of 801 nucleotides of EGFR affecting the external domain, known to be expressed in some human glioblastoma. As this mutation newly generates a glycine residue at the fusion point, a 14 amino acid peptide around the fusion junction including this glycine was chemically synthesised and used for immunisation of (B6 x DBA/2) F1 mice. Flow cytometric analysis showed 3C10 antibody staining of a mouse NIH/3T3 transfectant (ERM5) with the type III EGFR deletion-mutant gene, but not one with wild-type EGFR. The antibody immunoprecipitated the truncated EGFR protein with a molecular mass of approximately 140 kDa from ERM5 cells. Immunostaining of glioblastomas revealed binding in the case with the type III EGFR mutation, the five other specimens without the mutation being negative despite overexpression of EGFR in some cases.
一种小鼠单克隆抗体（IgG2b）3C10，是针对截短的表皮生长因子受体（EGFR）产生的，该受体由EGFR的801个核苷酸的（III型）框内缺失突变编码，影响外部结构域，已知在一些人胶质母细胞瘤中表达。由于这种突变在融合点新产生了一个甘氨酸残基，因此化学合成了围绕融合连接点的包括该甘氨酸的14个氨基酸的肽，并用于免疫（B6×DBA/2）F1小鼠。流式细胞术分析显示，3C10抗体可对携带III型EGFR缺失突变基因的小鼠NIH/3T3转染细胞（ERM5）进行染色，但对携带野生型EGFR的细胞则不能染色。该抗体从ERM5细胞中免疫沉淀出分子量约为140 kDa的截短EGFR蛋白。胶质母细胞瘤的免疫染色显示，在III型EGFR突变的病例中有结合，其他五个没有突变的标本尽管在某些情况下EGFR过表达，但结果为阴性。
