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Related Experiment Videos

Troybodies and pepbodies.

E Lunde1, V Lauvrak, I B Rasmussen

  • 1Division of Molecular Cell Biology, Department of Biology, P.O. Box 1050, Blindern, N-0316 Oslo, Norway.

Biochemical Society Transactions
|August 28, 2002
PubMed
Summary

Researchers engineered bacterial expression systems to create novel antibody fragments (pepbodies) with effector functions and developed Troybodies by inserting T-cell epitopes into antibodies to enhance T-cell activation for potential multi-vaccine applications.

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Area of Science:

  • Immunotechnology
  • Protein Engineering
  • Vaccine Development

Background:

  • Antibodies with effector functions are typically produced in mammalian cells, limiting bacterial production to non-effector fragments like scFv and Fab.
  • Current methods lack efficient bacterial production of functional antibody fragments and targeted T-cell activation strategies.

Purpose of the Study:

  • To develop novel antibody fragments (pepbodies) produced in bacteria that retain binding to antibody effector molecules and potentially initiate effector functions.
  • To engineer T-cell epitope-antibody fusions (Troybodies) for targeted delivery to antigen-presenting cells (APCs) and enhanced T-cell activation.

Main Methods:

  • Isolated peptides binding to antibody effector molecules and cloned them as fusions to scFv and Fab fragments in bacterial expression vectors.

Related Experiment Videos

  • Genetically inserted T-cell epitopes into antibody constant domains targeting APC surface molecules, creating Troybodies.
  • Investigated T-cell epitope loading onto MHC class II on APCs and presentation to T-cells.

    • Main Results:

    • Successfully generated targeting fragments (pepbodies) with the ability to bind antibody effector molecules, some initiating effector functions.
    • Demonstrated that T-cell epitopes inserted into antibodies are loaded onto MHC class II on APCs and presented to T-cells.
    • Achieved a significant increase (up to four orders of magnitude) in T-cell activation compared to synthetic peptides.

    Conclusions:

    • Bacterial production of functional antibody fragments (pepbodies) and targeted T-cell activation strategies (Troybodies) are feasible.
    • Troybodies enhance T-cell activation significantly, offering a promising platform for vaccine development.
    • Further research aims to identify all suitable Ig constant domain loops for T-cell epitope insertion to create a multi-vaccine.