| First Author | Fernando GJ | Year | 1999 |
| Journal | Clin Exp Immunol | Volume | 115 |
| Issue | 3 | Pages | 397-403 |
| PubMed ID | 10193409 | Mgi Jnum | J:53533 |
| Mgi Id | MGI:1332904 | Doi | 10.1046/j.1365-2249.1999.00813.x |
| Citation | Fernando GJ, et al. (1999) Expression, purification and immunological characterization of the transforming protein E7, from cervical cancer-associated human papillomavirus type 16. Clin Exp Immunol 115(3):397-403 |
| abstractText | E7 is the major oncogenic protein produced in cervical cancer-associated human papillomavirus type 16 (HPV16). This protein was expressed in Escherichia coli as a glutathione-S-transferase (GST) fusion protein. E7-enriched inclusion bodies were collected from bacterial lysates, were solubilized in 10 M urea, and the protein was purified using anion exchange column chromatography. After removal of endotoxin with serial Triton X-114 extractions, material of high purity (about 90%) was obtained, which is suitable for use in a human clinical trial. This material was immunogenic, and when used as a vaccine, protected mice against challenge with an HPV16 E7 DNA transfected tumour cell line. Based on this observation, the E7GST fusion protein is currently being used in a human clinical trial of a vaccine against HPV16-induced cervical cancer. This fusion protein could be cleaved with thrombin to remove the GST fusion part and further purified by preparative SDS gel electrophoresis to obtain free E7 with > 98% purity. |
