Characterization of scFv-421, a single-chain antibody targeted to p53.
A gene encoding a single-chain antibody (scFv) which specifically binds the tumor suppressor protein p53 has been constructed from RNA of hybridoma cells producing Pab 421. scFv-421 which was expressed and purified from bacteria specifically binds p53. scFv-421, as well as the previously described scFv-FRP5 and -R1R (1), were expressed intracellularly in mammalian cells and targeted to different subcellular locations, including the nucleus, cytoplasm, and endoplasmic reticulum (ER). High levels of all ER targeted scFv proteins, but not nuclear or cytoplasmic targeted proteins, were found in transfected COS-1 cells. In an attempt to stabilize the proteins, sequences encoding the mouse immunoglobin CK constant domain were added to each scFv construct. This led to a moderate increase in the cytoplasmic expression of scFv-FRP5.
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Friedrich Miescher Institute, Basel, Switzerland.
Hynes
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NE
D000595:Amino Acid Sequence; D000818:Animals; D019556:COS Cells; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006825:Hybridomas; D007128:Immunoglobulin Fragments / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D007135:Immunoglobulin Variable Region; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D057127:Single-Chain Antibodies; D013347:Subcellular Fractions / Q000378:metabolism; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000276:immunology*
D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D016159:Tumor Suppressor Protein p53; C104215:scFv 421