Wels W, Harwerth I M, Zwickl M, Hardman N, Groner B, Hynes N E
Friedrich Miescher Institute, Basel, Switzerland.
Biotechnology (N Y). 1992 Oct;10(10):1128-32. doi: 10.1038/nbt1092-1128.
We have constructed genes expressing single-chain antigen binding proteins (scFv) which recognize the human erbB-2 receptor. These genes encode the heavy and light chain variable domains of an erbB-2 receptor specific monoclonal antibody, MAb FRP5, connected by a peptide linker. In order to express a bifunctional molecule, a bacterial alkaline phosphatase gene was fused 3' to the scFv gene. The scFv(FRP5) and scFv(FRP5)-alkaline phosphatase fusion protein (scFv(FRP5)-PhoA) expressed in E. coli specifically recognize the human erbB-2 protein and compete with MAb FRP5 for binding to the receptor. The bound scFv(FRP5)-PhoA protein can be detected directly on tumor cells using a substrate for alkaline phosphatase, showing that the chimeric protein retains both binding and enzymatic activity.
我们构建了表达识别人类erbB-2受体的单链抗原结合蛋白（scFv）的基因。这些基因编码erbB-2受体特异性单克隆抗体MAb FRP5的重链和轻链可变结构域，它们通过一个肽接头相连。为了表达一种双功能分子，将细菌碱性磷酸酶基因融合到scFv基因的3'端。在大肠杆菌中表达的scFv(FRP5)和scFv(FRP5)-碱性磷酸酶融合蛋白（scFv(FRP5)-PhoA）能特异性识别人类erbB-2蛋白，并与MAb FRP5竞争结合该受体。使用碱性磷酸酶的底物可直接在肿瘤细胞上检测到结合的scFv(FRP5)-PhoA蛋白，这表明嵌合蛋白保留了结合活性和酶活性。
