Journal Article

Bioactive IL7-diphtheria fusion toxin secreted by mammalian cells

S. Shulga-Morskoy and B.E. Rich

in Protein Engineering, Design and Selection

Volume 18, issue 1, pages 25-31
Published in print January 2005 | ISSN: 1741-0126
Published online March 2005 | e-ISSN: 1741-0134 | DOI: https://dx.doi.org/10.1093/protein/gzi007

Show Summary Details

Preview

A number of targeted cytotoxic agents have been developed that selectively kill malignant or otherwise pathological cells. These engineered proteins consist of a potent cytotoxic element connected to a ligand domain that binds to specific molecules on the surface of the target cell. Several of these agents have shown promise in clinical trials and one is currently administered to patients. A significant technical obstacle that has impeded the development of some of these toxins is the difficulty of preparing certain recombinant proteins in properly folded forms. These fusion proteins have generally been produced in bacteria requiring them to be denatured and renatured in vitro. For some proteins this is an efficient process whereas for others it is not. We describe here a system to produce fusion toxins rapidly and efficiently by engineering mammalian cells to secrete them as properly folded molecules which can be purified in native form from cell culture medium. We have used this system to produce highly active preparations of DAB389-IL7, a molecule consisting of the catalytic and transmembrane domains of diphtheria toxin fused to interleukin 7. This system is generalizable and can be used to produce and evaluate rapidly fusion toxins incorporating novel or uncharacterized ligands.

Keywords: diphtheria toxin; fusion protein; interleukin-7; leukemia; lymphoma

Journal Article.  4896 words.  Illustrated.

Subjects: Proteins