Construction and expression of novel immunotoxin cpIL-4(13D)-PE38KDEL with increased activity |
| |
Authors: | J-X?Cui J-F?Ji A-G?Lv Email author" target="_blank">W-F?WuEmail author |
| |
Institution: | (1) Institute of Applied Ecology, Chinese Academy of Sciences, No. 72, Wenhua Road, 110016 Shenyang City, Liaoning Province, P. R. China;(2) Graduate School of the Chinese Academy of Sciences, 100039 Beijing, P. R. China;(3) Beckman Research Institute, City of Hope National Medical Center, 1500 Duarte Rd. Duarte, 91010, CA, USA |
| |
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. |
| |
Keywords: | immunotoxin interleukin-4 Pseudomonas exotoxin A |
本文献已被 PubMed SpringerLink 等数据库收录! |
|