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| Content Provider | Springer Nature Link |
|---|---|
| Author | Huang, Li Fen Liu, Yu Kuo Lu, Chung An Hsieh, Shie Liang Yu, Su May |
| Copyright Year | 2005 |
| Abstract | Human serum albumin (HSA) is the most widely used clinical serum protein. Currently, commercial HSA can only be obtained from human plasma, due to lack of commercially feasible recombinant protein expression systems. In this study, inducible expression and secretion of HSA by transformed rice suspension cell culture was established. Mature form of HSA was expressed under the control of the sucrose starvation-inducible rice α Amy3 promoter, and secretion of HSA into the culture medium was achieved by using the α Amy3 signal sequence. High concentrations of HSA were secreted into culture medium in a short time (2–4 days) by sucrose depletion after cell concentrations had reached a peak density in culture medium containing sucrose. The recombinant HSA had the same electrophoretic mobility as commercial HSA and was stable and free from apparent proteolysis in the culture medium. In a flask scale culture with repeated sucrose provision-depletion cycles, HSA was stably produced with yields up to 11.5% of total medium proteins or 15 mg/L per cycle after each sucrose provision-depletion cycle. A bubble column type bioreactor was designed for production of HSA. In the bioreactor scale culture, HSA was produced with yields up to 76.4 mg/L 4 days after sucrose depletion. HSA was purified from the culture medium to high purity by a simple purification scheme. Enrichment of HSA in culture medium simplifies downstream purification, minimizes protease degradation, and may reduce production cost. The combination of a DNA construct containing the α Amy3 promoter and signal sequence, and the use of a rice suspension cell culture can provide an effective system for the production of recombinant pharmaceutical proteins. |
| Starting Page | 569 |
| Ending Page | 581 |
| Page Count | 13 |
| File Format | |
| ISSN | 09628819 |
| Journal | Transgenic Research |
| Volume Number | 14 |
| Issue Number | 5 |
| e-ISSN | 15739368 |
| Language | English |
| Publisher | Kluwer Academic Publishers |
| Publisher Date | 2005-01-01 |
| Publisher Place | Dordrecht |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | α-amylase promoter human serum albumin rice suspension cell protein secretion sucrose starvation Human Genetics Plant Sciences Plant Genetics & Genomics Animal Genetics and Genomics |
| Content Type | Text |
| Resource Type | Article |
| Subject | Genetics Animal Science and Zoology Biotechnology Agronomy and Crop Science |
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