Expressing Intrabodies in Mammalian Cells

The intrabody approach is a gene-based technology developed to neutralize or modify the function of intracellular and extracellular target antigens. Since intrabodies can potentially target all different multiple isoforms of a protein, they can act as therapeutic molecules for the treatment of viral pathologies, cancer, and misfolding diseases. This chapter provides protocols for application of intrabodies in mammalian cells. We describe all the steps to express intrabodies in different subcellular compartments and to analyze the biochemical properties of scFv fragments and their intracellular distribution. We focus on protocols to study the antigen–intrabody complex in the secretory compartment and to analyze the effects of intrabody expression on membrane target protein.