Abstract
Interleukin-4 receptors (IL-4Rs) are expressed on a wide variety of human cancer cells, and therefore it may be a good option to treat IL-4R-bearing tumors with IL-4-fusing immunotoxins. In this study, the gene encoding human interleukin-4 mutein cpIL-4(13D) was obtained through overlapping polymerase chain reaction. A chimeric immunotoxin was constructed by genetically fusing the mutein cpIL-4(13D) to a modified version of Pseudomonas exotoxin A (PE38KDEL) and was expressed in Escherichia coli AD494 (DE3). The expression level of the fusion protein was about 30% of the total bacterial protein assessed by SDS-PAGE analysis. After purification by affinity chromatography and anion exchange chromatography, the chimeric protein was tested for its cytotoxicity. Our data show that cpIL-4(13D)-PE38KDEL has improved cytotoxicity on IL-4R-bearing tumor cells in comparison with other IL-4-fusing immunotoxins and might be useful in treating tumors with a large number of IL-4Rs.
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Translated from Biokhimiya, Vol. 70, No. 1, 2005, pp. 77–84. Original Russian Text Copyright © 2005 by Cui, Ji, Lv, Wu. Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM04-086, October 31, 2004.
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Cui, JX., Ji, JF., Lv, AG. et al. Construction and expression of novel immunotoxin cpIL-4(13D)-PE38KDEL with increased activity. Biochemistry (Moscow) 70, 62–68 (2005). https://doi.org/10.1007/PL00021758
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DOI: https://doi.org/10.1007/PL00021758