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Synthesis of an Intein-mediated Artificial Protein Hydrogel
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Generation of Potent Anti-HER1/2 Immunotoxins by Protein Ligation Using Split Inteins

Thomas Pirzer1, Kira-Sophie Becher2, Marcel Rieker3,4

  • 1Institute for Organic Chemistry and Biochemistry , Technische Universität Darmstadt , Alarich-Weiss-Strasse 4 , D-64287 Darmstadt , Germany.

ACS Chemical Biology
|June 20, 2018
PubMed

Insights

Researchers developed a novel method to create potent antibody immunotoxins for cancer therapy. This technique uses split inteins for efficient production of full-length antibody toxins, enhancing therapeutic potential.

Area of Science:

  • Biotechnology
  • Molecular Biology
  • Cancer Therapeutics

Background:

  • Cell-targeting protein toxins are promising for cancer therapy, aiming to improve efficacy and reduce side effects.
  • Current methods often use antibody fragments produced in bacteria, which may lack optimal properties like longer circulation times.
  • Producing full-length antibody immunotoxins is challenging due to toxicity in eukaryotic hosts.

Purpose of the Study:

  • To develop a method for producing full-length antibody immunotoxins using self-splicing split inteins.
  • To overcome limitations of current immunotoxin production methods.
  • To generate specific and potent immunotoxins for potential cancer treatment.

Main Methods:

  • Recombinant fusion of an N-terminal intein fragment to antibody heavy chains (trastuzumab, HER2-targeting; nanobody-Fc, HER1-targeting).
  • Expression of antibody fragments in Expi293F cells and toxin fragments (fused to maltose binding protein) in E. coli.
  • In vitro splicing of intein fragments to assemble full-length antibody immunotoxins.

Main Results:

  • Successfully produced full-length antibody immunotoxins targeting HER2 and HER1 receptors.
  • Achieved specific and potent immunotoxin activity.
  • Generated immunotoxins with IC50 values in the mid- to subpicomolar range.

Conclusions:

  • Self-splicing split inteins enable efficient production of full-length antibody immunotoxins.
  • This method overcomes toxicity issues associated with eukaryotic expression of immunotoxins.
  • The generated immunotoxins show high potency and specificity, offering a promising approach for cancer therapy.

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