Chemical engineering of cell penetrating antibodies

J Immunol Methods. 2001 Aug 1;254(1-2):137-45. doi: 10.1016/s0022-1759(01)00410-0.

Abstract

Antibodies, being exquisitely specific tools in biology, are routinely used to detect and identify intra-cellular structures. However, current intra-cellular application of antibodies requires that the membrane be rendered leaky, resulting in the death of cells. Here, we present a novel method to allow antibodies to penetrate the cellular membrane of living cells without affecting cell viability. A peptide (MTS, membrane transport sequence) that facilitates transport across membranes has been site-specifically attached to antibodies. MTS-antibodies enter the living cells in culture and can be detected by immunofluorescence and ELISA after extraction. Cellular structures are visualized in living cells using a specific MTS-antibody. Antibodies with membrane penetrating properties can become an important tool for the study of intra-cellular processes in living cells. Furthermore, such membrane penetrating antibodies can be used to selectively stimulate or suppress functions of the cellular machinery.

MeSH terms

  • 3T3 Cells
  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / metabolism*
  • Antibodies, Neoplasm / metabolism*
  • Cross-Linking Reagents
  • Cytoskeleton
  • Fibroblast Growth Factor 4
  • Fibroblast Growth Factors / metabolism*
  • Fluorescence
  • Humans
  • Lymphoma, B-Cell / immunology
  • Mice
  • Molecular Sequence Data
  • Protein Engineering
  • Protein Sorting Signals*
  • Protein Transport
  • Proto-Oncogene Proteins / metabolism*
  • Staining and Labeling / methods

Substances

  • Antibodies, Monoclonal
  • Antibodies, Neoplasm
  • Cross-Linking Reagents
  • FGF4 protein, human
  • Fgf4 protein, mouse
  • Fibroblast Growth Factor 4
  • Protein Sorting Signals
  • Proto-Oncogene Proteins
  • Fibroblast Growth Factors