Abstract
Preparation and Evaluation of activity of Highly-soluble Anticancer Protein Drug TATm-survivin (T34A)
Department of Biochemistry, Weihai Vocational College, Weihai City Science and Technology Park, 264210, Weihai, Shandong, China
Correspondence Address:
Department of Biochemistry, Weihai Vocational College, Weihai City Science and Technology Park, 264210, Weihai, Shandong, China, E-mail: haiyi_zhang@163.com
TATm-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 TATm-survivin (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 TATm-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 TATm-survivin (T34A) was obtained with a purity of 96 %, which inhibited the proliferation of human pancreas carcinoma cell lines SW1990. In comparison to denatured TATm-survivin (T34A), soluble TATm-survivin (T34A) did not exhibit higher bioactivity as expected. This study represented a novel strategy to obtain highly soluble form of TATm-survivin (T34A) and would provide useful information for production of other soluble tat-mediated fusion proteins.
