| 摘要 |
In order to activate gene technologically produced, heterologous, eucaryotic proteins containing disulphide bridges, after expression in procaryotes by cell decomposition, solubilizing under denaturing and reducing conditions and activating under oxidizing conditions in the presence of reduced glutathione/glutathione disulphide (GSH/GSSG), one operates either in the activating stage at a pH value of 9 to 12, a reduced glutathione (GSH) concentration of 0.1 to 20 mmol/l, an oxidized glutathione (GSSG) concentration of 0.01 to 3 mmol/l, and with a nondenaturing concentration of the denaturing agent, or by separating the reducing/denaturing agent, the thiol group of the protein being transferred into the mixed disulphides of protein and glutathione by the addition of GSSG under denaturing conditions, and a GSH concentration of 0.5 to 5 mmol/l and a non-denaturing concentration of the denaturing agent being set up in the activating stage at a pH value of 7 to 10.5; the method is particularly valuable for t-PA and interferon.
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