| Journal: | Brazilian Journal of Pharmaceutical Sciences |
| Database: | PERIÓDICA |
| System number: | 000451455 |
| ISSN: | 1984-8250 |
| Authors: | Farshdari, Farzaneh1 Ahmadzadeh, Maryam2 Nematollahi, Leila3 Mohit, Elham2 |
| Institutions: | 1Islamic Azad University, Faculty of Advanced Sciences & Technology, , Teherán. Irán 2Shahid Beheshti University of Medical Sciences, School of Pharmacy, Teherán. Irán 3Pasteur Institute of Iran, Biotechnology Research Center, Teherán. Irán |
| Year: | 2020 |
| Volumen: | 56 |
| Country: | Brasil |
| Language: | Inglés |
| Document type: | Artículo |
| Approach: | Experimental, aplicado |
| English abstract | The relationship between the expression of HER2 and malignity of breast tumors has led to the generation of antibodies targeting HER2+ tumors. In addition, the expression of scFvs, as the smallest antigen-binding region of antibody containing two disulfide bonds in Escherichia coli often results in accumulating non-functional protein in the cytoplasm. A redox-modified strain of E. coli such as Origami (DE3) may facilitate the formation of proper disulfide bond in cytoplasm. The present study aimed to optimize the expression of anti-HER2 scFv in Origami and evaluate the influence of induction temperature, and host strain on the solubility of the protein. To this aim, chemically synthesized anti-HER2 scFv of Trastuzumab was cloned in pET-22b (+). The results demonstrated that anti-HER2 scFv is expressed in Origami, purified by using Ni-NTA column, and detected by anti-His antibody in Western blot analysis. The highest anti-HER2 scFv expression in Origami was achieved 24 h after IPTG induction (1 mM) at 37 ºC. Further, the total anti-HER2 scFv expression level was higher in BL21, compared to Origami strain. However, the ratio of soluble/insoluble forms of anti-HER2 scFv increased in Origami strain. Furthermore, higher soluble expression was achieved when the culture of recombinant Origami was conducted at lower temperature (25 ºC) |
| Disciplines: | Medicina |
| Keyword: | Oncología, Medicina experimental, Genética, Cáncer, Mama, Regulación génica, HER2, Sobreexpresión génica, Anticuerpos, Blancos terapéuticos |
| Keyword: | Oncology, Experimental medicine, Genetics, Cancer, Breast, Gene regulation, HER2, Gene overexpression, Antibodies, Therapeutic targets |
| Full text: | Texto completo (Ver HTML) Texto completo (Ver PDF) |
