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- Title
Optimization of a recombinant BlaR-CTD protein formulation using the response surface methodology.
- Authors
Abdolvahab, Mohadeseh Haji; Safari, Mojdeh; Hasannejad, Farkhonde; Asefi, Nika; Salimi, Alireza; Nazari, Mahboobeh
- Abstract
The sequence of a carboxy-terminal of the β-lactam sensor-transducer protein (BlaR-CTD) from Bacillus licheniformis ATCC14580 was extracted from US7745193B2 patent and expressed in E. coli using pColdI vector as a soluble His-tag recombinant protein. In this study, several excipients were used to improve the stability of recombinant BlaR-CTD and obtain the optimal formulation for this protein using response surface methodology (RSM)/ Central Composite Design (CCD). Total protein concentration was measured by UV spectroscopy and the Bradford test. A total of 7 various factors were designed using four different excipients including Glycerol, Sucrose, Triton x-100, and Tween-20, and three different buffers like Tris, Borate, and PBS. By obtaining suitable excipients and buffer i.e. glycerol and sucrose, pH ranging from 7 to 9 were evaluated. The pH 7.62, glycerol 15.35%, and sucrose 152.52 mM were determined as the most suitable for improving the thermal stability of recombinant BlaR-CTD.
- Subjects
RESPONSE surfaces (Statistics); ESCHERICHIA coli; BACILLUS licheniformis; TRITON X-100; SUCROSE; PYRAZOLYL compounds; THERMAL stability; RECOMBINANT proteins
- Publication
Journal of Biological Engineering, 2024, Vol 18, Issue 1, p1
- ISSN
1754-1611
- Publication type
Article
- DOI
10.1186/s13036-023-00399-9