Intracellular expression of the anti-erbB-2 sFv N29 fails to accomplish efficient target modulation

Academic Article

Abstract

  • The use of intracellular single chain antibodies has recently emerged as a highly efficient method of downregulating or ablating protein expression. In this regard, we have demonstrated that a single chain antibody directed against the extracellular domain of the erbB-2 molecule causes a specific toxicity in erbB-2 positive tumor types. To further investigate the mechanism of this effect, we developed a second anti-erbB-2 sFv predicted to recognize an alternate extracellular epitope of the erbB-2 molecule. When produced as a secreted protein from the erbB-2 negative COS-1 cell line, this sFv binds specifically to erbB-2 positive cells, indicating that cellular machinery is able to produce a properly folded and functional sFv protein. However, by several assays, this sFv was shown to be unable to retain the erbB-2 protein within the ER. These negative results have implications for the evaluation and utilization of sFv knockout strategies in experimental contexts.
  • Digital Object Identifier (doi)

    Pubmed Id

  • 25251366
  • Author List

  • Grim JE; Siegal GP; Alvarez RD; Curiel DT
  • Start Page

  • 699
  • End Page

  • 703
  • Volume

  • 250
  • Issue

  • 3