Generation of Murine scFv Intrabodies from B-Cell Hybridomas

The intracellular expression of recombinant single-chain Fv (scFv) molecules—termed intrabodies—has potential advantages for therapeutic use (1 ,2 ). These single polypeptide chains are made of one heavy-chain variable region (VH) linked through a flexible spacer, usually a repeated motif of 3� GGGGS (although some others have also been described) (3 ), to one light chain variable region (VL). The VH-linker-VL sequence can be inverted with no loss of the binding. Shortened linkers have been used to produce scFvs of divalent or bispecific forms (3 ,4 ).