Design of a modular immunotoxin connected by polyionic adapter peptides

J Mol Biol. 2003 Mar 21;327(2):445-52. doi: 10.1016/s0022-2836(03)00141-4.

Abstract

Immunotoxins are genetically engineered fusion proteins of an antibody Fv fragment and a toxin from bacteria or plants, which function as anti-cancer therapeutics. Here, we describe a new generation of immunotoxins in which both proteins do not form a single fusion protein but are coupled specifically via cysteine-containing polyionic fusion peptides. The engineered Pseudomonas exotoxin PE38 was N-terminally fused to the peptide E(8)C. In combination with the disulfide-stabilized Fv fragment of the tumor-specific antibody B3, which was extended by the peptide R(8)CP, the fusion peptides ensured a specific and covalent coupling of the Fv fragment and the toxin. The resulting immunotoxin was as active and as specific as an immunotoxin consisting of a fusion protein of the same antibody fragment connected to the toxin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ADP Ribose Transferases / chemistry*
  • ADP Ribose Transferases / genetics
  • Antibodies, Monoclonal / genetics
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / metabolism
  • Bacterial Toxins / chemistry*
  • Bacterial Toxins / genetics
  • Cell Death / drug effects
  • Disulfides / chemistry*
  • Drug Design*
  • Exotoxins / chemistry*
  • Exotoxins / genetics
  • Immunoglobulin Fragments / chemistry
  • Immunoglobulin Fragments / genetics
  • Immunoglobulin Fragments / metabolism
  • Immunotoxins / chemistry*
  • Immunotoxins / genetics
  • Immunotoxins / pharmacology*
  • Models, Molecular
  • Neoplasm Proteins / genetics*
  • Neoplasm Proteins / immunology
  • Neoplasm Proteins / metabolism
  • Peptide Fragments / chemistry
  • Peptide Fragments / metabolism*
  • Plasmids
  • Pseudomonas aeruginosa
  • Pseudomonas aeruginosa Exotoxin A
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / pharmacokinetics
  • Recombinant Fusion Proteins / pharmacology
  • Structure-Activity Relationship
  • Virulence Factors / chemistry*
  • Virulence Factors / genetics

Substances

  • Antibodies, Monoclonal
  • Bacterial Toxins
  • Disulfides
  • Exotoxins
  • Immunoglobulin Fragments
  • Immunotoxins
  • Neoplasm Proteins
  • Peptide Fragments
  • Recombinant Fusion Proteins
  • Virulence Factors
  • immunoglobulin Fv
  • immunotoxin LMB-7
  • ADP Ribose Transferases