Preparation and Evaluation of activity of Highly-soluble Anticancer Protein Drug TATm-survivin (T34A)

Abstract

Zhang et al.: Preparation of Soluble Protein Drug T34ATAT m -survivin (T34A), an anticancer protein drug, displayed pro-apoptotic bioactivity against various cancer cells. It was expressed in Escherichia coli as inclusion bodies. To test the bioactivity of soluble TAT msurvivin (T34A), a feasible strategy was first developed for the soluble expression and purification. Effect of zinc ion and induction temperature was investigated to improve the solubility and overall production level of TAT m -survivin (T34A). High solubility (92 %) was achieved by addition of zinc ion and temperature downshift after induction. An efficient protocol of purification was established by heat release, ammonium sulphate precipitation, anion exchange and heparin affinity chromatography. Purified TAT m -survivin (T34A) was obtained with a purity of 96 %, which inhibited the proliferation of human pancreas carcinoma cell lines SW1990. In comparison to denatured TAT m -survivin (T34A), soluble TAT m -survivin (T34A) did not exhibit higher bioactivity as expected. This study represented a novel strategy to obtain highly soluble form of TAT m -survivin (T34A) and would provide useful information for production of other soluble tat-mediated fusion proteins.