Intracellular expression of single chain antibodies reverts ErbB-2 transformation

摘要: We report a novel approach for specific in vivo inactivation of the ErbB-2 receptor tyrosine kinase and suppression ErbB-2-induced transformation. Genes encoding single chain antibodies that specifically bind to extracellular domain human were constructed expressed intracellularly NIH/3T3 fibroblasts transformed by activated ErbB-2. The are derived from monoclonal FRP5 FWP51 (Harwerth, I.M., Wels, W., Marte, B. M., Hynes, N. E. (1992) J. Biol. Chem. 267, 15160-15167) composed heavy light variable domains connected flexible peptide linker. provided with: 1) an N-terminal hydrophobic leader sequence target their synthesis lumen endoplasmic reticulum, 2) C-terminal retention signal prevent secretion. When ErbB-2-transformed cells, bound prevented its transit through reticulum. This resulted functional reversion phenotype. is first demonstration targeted stable cellular oncoprotein via intracellular antibody expression. use such strategy represents simple powerful study function receptors other proteins.