Woodchuck Hepatitis Virus Post-Transcriptional Regulation Element (WPRE) Promotes Anti-CD19 BiTE Expression in Expi293 Cells     

Reza Moazzami  Hasan Mirzahoseini  Leila Nematollahi  Farzaneh Barkhordari  Mozhgan Raigani  Fatemeh Hajari Taheri  Fereidoun Mahboudi  Fatemeh Davami 《Iranian Biomedical Journal》2021,25(4):275

Background:Bispecific antibodies represent an important class of mAbs, with great therapeutic potentials due to their ability to target simultaneously two distinct epitopes. The generation of functional bispecific antibodies with the highest possible yields is particularly critical for the production of these compounds on industrial scales. Anti- CD3 × CD19 bsAb is a bispecific T-cell engager (BiTE) currently used for treating ALL. Herein, we have tried to optimize the expression level of this antibody in mammalian hosts. Methods:WPRE sequence was incorporated at the 3’ end of the expression cassette. This modification resulted in a notable about two-fold increase in the expression of the bsAb in the Expi293 cell line. Results & Conclusion:Follow-up flow cytometry analysis demonstrated the binding properties of the produced antibody at acceptable levels, and in vitro bioactivity assays showed that this product is potent enough for targeting and destroying CD19-positive cells. Our findings show that WPRE enhances the expression of this type of bispecific mAbs in HEK-293 family cell lines. This approach can be used in biopharma industry for the mass production of anti-CD3 × CD19 bispecific antibody. Key Words: Acute lymphoblastic leukemia, Bispecific antibodies, Monoclonal antibody  相似文献   

The effect of form and level of inorganic N on nitrogen use efficiency of sugarcane grown in pots     

Elliosha Hajari  Sandy J. Snyman 《Journal of plant nutrition》2017,40(2):248-257

To-date, assessments of nitrogen use efficiency (NUE) of sugarcane have not included the contribution of its components, nitrogen uptake efficiency (NUpE) and nitrogen utilization efficiency (NUtE). This study determined these values, based on biomass and plant nitrogen (N) content, in two four-month-old pot-grown genotypes. The treatments included six N regimes, with nitrate (NO₃^—N) or ammonium (NH₄⁺-N) supplied alone, or as NO₃^?-N for the first 6 weeks and then NH₄⁺-N until harvest, each as 4 or 20 mM. Regardless of the N form, NUE was higher at four than at 20 mM due to significantly higher NUpE at low N supply. The results indicated that there was luxury N uptake and preference for NH₄⁺-N nutrition, which resulted in the highest determined NUE. There were significant differences between genotypes in biomass, morphological growth parameters, N uptake, total plant N and NUE, the latter matching previously established sucrose yield-based NUE field rankings.  相似文献