High Expression of a cDNA Coding for Human Lymphotoxin alpha Derivative in E. coli and Purification of Its Protein Product.
1998
Using PCR, a cDNA coding for human lymphotoxin α derivative (hLTαDa) lacking 27 amino acids at Nterminal of natural hLT was constructed. The expression construct was expressed in E.coli BL21(DE3). The product of expression was in the form of inclusion bodies, and accounted for 60%-80% of total bacterial proteins . The product protein was purified to over 95% by treatments of inclusion bodies. The specific activity of hLTαDa was above 2×108 IU(per mg protein). Its cytotoxic activity can be neutralized by monoclone antibody against hLT. The antitumor effects of hLTαDa were also tested in vitro and in vivo.
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