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| Content Provider | Springer Nature Link |
|---|---|
| Author | Zhong, Genshen Yu, Aiping Shi, Bingxing Liu, Yang Wu, Chutse |
| Copyright Year | 2008 |
| Abstract | SFH, a recombinant staphylokinase-based fusion protein linked by the factor Xa recognition peptide at the N-terminus of hirudin, is a promising therapeutic candidate for thromboembolic diseases. To develop SFH into a new thrombolytic agent, scaled-up production was carried out to provide sufficient preparation for animal safety and clinical studies. Here, we describe a pilot-scale cultivation and purification process for the production of SFH. A high-cell-density fed-batch cultivation for the production of SFH in E. coli was developed in a 40-L bioreactor, which produced about 1.1 g/L of recombinant protein. SFH was purified to homogeneity from the E. coli lysate by expanded bed adsorption chromatography and anion-exchange chromatography, with over 99% purity and 54% recovery. Moreover, the residual endotoxin content was less than 0.5 EU/mL. The molecular weight and in vitro bioactivity of SFH were also determined by electrospray ionization-mass spectrometry (ESI-MS) and fibrinolytic activity assay, respectively. |
| Starting Page | 75 |
| Ending Page | 81 |
| Page Count | 7 |
| File Format | |
| ISSN | 16733509 |
| Journal | Frontiers in Biology |
| Volume Number | 4 |
| Issue Number | 1 |
| e-ISSN | 16733622 |
| Language | English |
| Publisher | SP Higher Education Press |
| Publisher Date | 2008-08-15 |
| Publisher Institution | Chinese Universities |
| Publisher Place | Heidelberg |
| Access Restriction | Subscribed |
| Subject Keyword | staphylokinase E. coli purification endotoxin Life Sciences |
| Content Type | Text |
| Resource Type | Article |
| Subject | Ecology Genetics Ecology, Evolution, Behavior and Systematics Biotechnology |
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