Expression and Function Test of Hemoglobin Beta-alpha1 Microglobulin Fusion Protein
| Autor: | Wu, Guan-Wei, 吳冠緯 |
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| Rok vydání: | 2019 |
| Druh dokumentu: | 學位論文 ; thesis |
| Popis: | 107 In response to global blood bank shortage and prevention of blood-borne diseases, scientists have been studying blood substitutes such as perfluorocarbon and hemoglobin-based oxygen carriers (HBOC). Chemically modified or genetically recombined hemoglobin (Hb) has resolved the cause of kidney failure through inceasing the size of Hb and prolonged the half-life of Hb in plasma, However, the side effects of increased blood pressure and oxidative stress have not been fully resolved in the clinical trials. In this study, α-1-microglobulin (A1M) was fused with Hbβ for scavenging O2- and restoring oxyHbβ from metHb. NADH/NADPH and vitamin C can be used by A1M as electron donors to accelerate the reduction of metHb. The Hb proteins were expressed in the transformed cells, by observing the pink color of the cells, and the SDS-PAGE protein electrophoresis, The structures of the proteins become unstable during the process of purification and concentration, resulting in the heme released from Hb. Various strategies of cell disruption and protein purification have been developed to stabilize the proteins, but they were still not stable enough to pursue the functional studies. |
| Databáze: | Networked Digital Library of Theses & Dissertations |
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