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Intracellular Immunization with Cytosolic Recombinant Antibodies

Bio/Technology volume 12, pages 396–399 (1994)Cite this article

Abstract

We report the application of a strategy to inactivate cellular proteins in vertebrate cells based on the intracellular expression of immunoglobulin genes. We have selected, in this instance, the p21 protein, encoded by the ras protooncogene, as a target protein. The variable regions of the neutralizing anti-p21ras monoclonal antibody Y13-259 were cloned in vectors for the expression of either the whole antibody molecule or its singlechain Fv fragment (ScFv) derivative. In order to target the recombinant antibodies to the cytosol, their hydrophobic leader sequence for secretion was mutated or deleted. When these proteins are expressed in the cytosol of Xenopus laevis oocytes they colocalize with the endogenous p21ras protein in the cytoplasmic face of the oocyte plasma membrane, and they markedly inhibit the H1 kinase activity induced by insulin. Moreover, cytosolic anti-p21ras ScFv fragments block the ensuing meiotic maturation. Thus the intracellular expression of both whole antibodies and antibody domains can be used to block a biological function.

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Author notes

  1. Silvia Biocca and Antonino Cattaneo: Corresponding authors at: Institute of Neurobiology, Viale K. Marx 15, 00137 Roma, Italy.

Authors and Affiliations

  1. Institute of Neurobiology, Viale K. Marx 15, 00137, Roma, Italy

    Silvia Biocca & Antonino Cattaneo

  2. Institute of Cell Biology, CNR, Viale K. Marx 15, 00137, Roma, Italy

    Paola Pierandrei-Amaldi & Nadia Campioni

  3. International School for Advanced Studies (SISSA), Trieste, Italy

    Antonino Cattaneo

Authors
  1. Silvia Biocca
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  2. Paola Pierandrei-Amaldi
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  3. Nadia Campioni
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  4. Antonino Cattaneo
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Biocca, S., Pierandrei-Amaldi, P., Campioni, N. et al. Intracellular Immunization with Cytosolic Recombinant Antibodies. Nat Biotechnol 12, 396–399 (1994). https://doi.org/10.1038/nbt0494-396

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