T cell-targeted immunofusion proteins from Escherichia coli.

The Journal of Biological Chemistry
M BetterD M Fishwild

Abstract

Fusion proteins between cell-targeting domains and cytotoxic proteins should be particularly effective therapeutic reagents. We constructed a family of immunofusion proteins linking humanized Fab, F(ab')2, or single chain antibody forms of the H65 antibody (which recognizes the CD5 antigen on the surface of human T cells) with the plant ribosome-inactivating protein gelonin. We reasoned that such an immunofusion would kill human target cells as efficiently as the previously described chemical conjugates of H65 and gelonin (Better M., Bernhard, S. L., Fishwild, D. M., Nolan, P. A., Bauer, R. J., Kung, A. H. C., and Carroll, S. F. (1994) J. Biol. Chem. 269, 9644-9650) if both the recognition and catalytic domains remained active, and a proper linkage between domains could be found. Immunofusion proteins were produced in Escherichia coli as secreted proteins and were recovered directly from the bacterial culture supernatant in an active form. All of the immunofusion proteins were purified by a common process and were tested for cytotoxicity toward antigen-positive human cells. A 20-60-fold range of cytotoxic activity was seen among the fusion family members, and several fusion proteins were identified which are approximately as ac...Continue Reading

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Citations

Dec 24, 1997·Leukemia Research·R J Kreitman
Oct 18, 2000·Biochemical and Biophysical Research Communications·A SchmidtH Zinke
Mar 21, 2012·Expert Opinion on Drug Metabolism & Toxicology·Xiaoying ChenWei-Chiang Shen
Jan 16, 2007·Protein Expression and Purification·Jerica SaboticJoze Brzin
Jul 2, 2003·The Journal of Biological Chemistry·Chester J ProvodaKyung-Dall Lee
Oct 10, 2001·The Journal of Immunology : Official Journal of the American Association of Immunologists·P L TazzariF Stirpe

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