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|Title:||Role of polyethyleneimine in the purification of recombinant human tumour necrosis factor beta|
|Authors:||Loh, K.C. |
Tumor necrosis factor
|Citation:||Loh, K.C., Yao, Z.J., Yap, M.G.S., Chung, M.C.M. (1997-01-31). Role of polyethyleneimine in the purification of recombinant human tumour necrosis factor beta. Journal of Chromatography A 760 (2) : 165-171. ScholarBank@NUS Repository. https://doi.org/10.1016/S0021-9673(96)00779-0|
|Abstract:||The chromatographic behaviour of recombinant human tumour necrosis factor beta (rhTNF-β) (pI ~9.0) during cation-exchange chromatography at pH 7.5 is investigated. Without prior treatment of the Escherichia coli cell extract with polyethyleneimine (PEI), very little rhTNF-β was bound to the column. However, upon addition of 5% PEI (100 μl ml-1) to the cell lysate, rhTNF-β was shown to bind to cation-exchange columns normally. TNF-β was readily precipitated from the clarified cell extract by 20% ammonium sulphate, but only ca. 25% of this precipitate could be re-solubilized for further purification. However, when 5% PEI was included in the solubilization buffer, the balance of the rhTNF-β could be recovered. It is proposed that charge interaction between rhTNF-β and nucleic acids in the cell extract is responsible for both of these anomalous phenomena, and that PEI (a cationic polyelectrolyte) was able to disrupt this interaction by displacing rhTNF-β from the charge complex.|
|Source Title:||Journal of Chromatography A|
|Appears in Collections:||Staff Publications|
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