| 摘要 |
PURPOSE:To remove foreign proteins and pyrogenic substances and obtain purified product, by adsorbing a precursor of a refolded urokinase-like enzyme originated from microbial cells in a cation exchange resin and fractionally eluting the adsorbed substances. CONSTITUTION:A gene of a precursor (proUK) of a urokinase-like enzyme is integrated into an E.coli and the proUK is produced by the recombinant E.coli. The microbial cells are disintegrated and filtered, etc., to obtain an insoluble fraction, from which proUK is extracted with an alkaline aqueous solution. A refolded proUK is produced by controlling the pH of the extract to 8-10. The solution is directly adsorbed to a cation exchange resin and fractionally eluted to remove foreign proteins and obtain purified product. When the raw material solution contains salts at a concentration of e.g., >=50mM, the solution is preferably desalted e.g., by dialysis before passing through a column packed with a cation exchange resin. |
