HUMANIZATION OF AN ANTI-P185HER2 ANTIBODY FOR HUMAN CANCER-THERAPY
HUMANIZATION OF AN ANTI-P185HER2 ANTIBODY FOR HUMAN CANCER-THERAPY
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DOI：
10.1073/pnas.89.10.4285
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发表时间：
1992-05-15
影响因子：
11.1
通讯作者：
SHEPARD, HM
中科院分区：
文献类型：
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作者：
CARTER, P;PRESTA, L;SHEPARD, HM
针对人表皮生长因子受体2（p185 HER 2）的鼠单克隆抗体mumAb 4D 5特异性抑制过表达p185 HER 2的人肿瘤细胞的增殖。然而，mumAb 4D 5在人类癌症治疗中的功效可能受到人类抗小鼠抗体应答和缺乏效应子功能的限制。构建了“人源化”抗体humAb 4D 5 -1，其仅含有来自mumAb 4D 5的抗原结合环和人可变区框架残基加IgG 1恒定结构域。轻链和重链可变区分别使用311-mer和361-mer预组装的寡核苷酸通过“基因转化诱变”在一个步骤中同时人源化。humAb 4D 5 -1变体不阻断过表达p185 HER 2的人乳腺癌SK-BR-3细胞的增殖，尽管抗原结合紧密（K（d）= 25 nM）。通过分子建模设计的七种另外的人源化变体之一（humAb 4D 5 -8）分别比humAb 4D 5 -1和mumAb 4D 5更紧密地结合p185 HER 2抗原250倍和3倍。此外，humAb 4D 5 -8在阻断SK-BR-3细胞增殖方面具有与鼠抗体相当的效力。此外，humAb 4D 5 -8在支持针对SK-BR-3细胞的抗体依赖性细胞毒性方面比mumAb 4D 5有效得多，但它不能有效地杀死以较低水平表达p185 HER 2的WI-38细胞。
The murine monoclonal antibody mumAb4D5, directed against human epidermal growth factor receptor 2 (p185HER2), specifically inhibits proliferation of human tumor cells overexpressing p185HER2. However, the efficacy of mumAb4D5 in human cancer therapy is likely to be limited by a human anti-mouse antibody response and lack of effector functions. A "humanized" antibody, humAb4D5-1, containing only the antigen binding loops from mumAb4D5 and human variable region framework residues plus IgG1 constant domains was constructed. Light- and heavy-chain variable regions were simultaneously humanized in one step by "gene conversion mutagenesis" using 311-mer and 361-mer preassembled oligonucleotides, respectively. The humAb4D5-1 variant does not block the proliferation of human breast carcinoma SK-BR-3 cells, which overexpress p185HER2, despite tight antigen binding (K(d) = 25 nM). One of seven additional humanized variants designed by molecular modeling (humAb4D5-8) binds the p185HER2 antigen 250-fold and 3-fold more tightly than humAb4D5-1 and mumAb4D5, respectively. In addition, humAb4D5-8 has potency comparable to the murine antibody in blocking SK-BR-3 cell proliferation. Furthermore, humAb4D5-8 is much more efficient in supporting antibody-dependent cellular cytotoxicity against SK-BR-3 cells than mumAb4D5, but it does not efficiently kill WI-38 cells, which express p185HER2 at lower levels.