چاپ صفحه |  صفحه نخست سامانه |  چکیده مقاله |  نویسندگان |  دانلود مقاله | دانشگاه علوم پزشکی تبریز |
| Abstract Single-chain variable fragments (scFvs) have gained increased attention among researchers in both academic and industrial felds owing to simple production in E. coli. The E. coli periplasm has been the site of choice for the expression of scFv molecules due to its oxidizing milieu facilitating correctly formation of disulfde bonds. Hence, the recovery of high-yield and biologically active species from the periplasmic space is a critical step at beginning of downstream processing. TES (Tris/EDTA/Sucrose) as a simple and efcient extraction method has been frequently used but under varied extraction conditions, over literature. This study, for the frst time, aimed to interrogate the efects of four independent variables (i.e., Tris–HCl concentration, bufer’s pH, EDTA concentration, and incubation time) and their potential interactions on the functional extraction yield of an scFv antibody from the periplasmic space of E. coli. The results indicated that the Tris–HCl concentration and pH are the most signifcant variables in the TES method and displayed a positive efect at their lower values on the functional extraction yield. Besides, the statistical analysis revealed 4 signifcant interactions between diferent variables. Here is the frst report on the successful application of a design of experiment based on a central composite design to establish a generic and optimal TES extraction condition. Accordingly, an optimal condition for TES extraction of scFv molecules from the periplasm of HB2151 at the exponential phase was developed as follows: 50 mM Tris–HCl at pH 7.2, 0.53 mM EDTA, and an incubation time of 60 min. |
| نویسنده | نفر چندم مقاله |
|---|---|
| محمد رضا توحیدکیا | دوم |
| بهزاد برادران | هفتم |
| امیر علی مختارزاده | ششم |
| طیبه مهدی پور | پنجم |
| نام فایل | تاریخ درج فایل | اندازه فایل | دانلود |
|---|---|---|---|
| s13568-020-01063-x.pdf | 1399/05/11 | 3974975 | دانلود |
