The Investigation Of The Separation And Purification Procedure Of Recombinant Human Granulocyte Colony-stimulating Factor

Recombinant human granulocyte colony-stimulating factor (rhG-CSF)injection is a liquid preparation prepared from recombinant proteins expressed in E.coli containing recombinant plasmids of the rhG-CSF gene. The recombinant proteins are isolated and purified after fermentation of the transformed E.coli.The US FDA authorized the sell of GCSF of Amgen Co. Ltd in June of 1996. Since the frist civil leucocyte promotion drug—Jilifen(rhG-CSF)—was authorized for production in October of 1996, many similar drugs were recommended. The protein of rhG-CSF was mostly purified as the form of inclusion body by civil corporations and minor was purified by secretion form. In our experiment, we broke down the cell wall by recursive freeze and thaw. The target protein was released from the bacteria in the low osmotic form. The protein solution included many impurities except the target protein after centrifuge. In advantage of the solubility difference, we obtained the target protein by twice using salt out and primarily eliminated the impurities. Then we attained the acquired purity of rhG-CSF by using different chromatography. The research was started from lyses and salt out, explored a better way of GCSF purify and chromatography. We obtained the high purity and recovery of GCSF by using the least steps and extremely low the cost of production.