Yields Biologically Active Protein with a Trimeric Structure That Binds to Both Tumor Necrosis Factor Receptors*
نویسندگان
چکیده
A fast and efficient method for medium scale purification of recombinant human tumor necrosis factor @ (rTNF-8) from Escherichia coli cells is described. The purified rTNF-@ displayed biological activity similar to rTNF-a in a WEHI 164 cell cytotoxicity assay. The titration curve of rTNF-8 and elution profiles of rTNFf l in gel filtration experiments were different from those of rTNF-a. However, light scattering and ultracentrifugation studies showed that both cytokines have trimeric structures in solution at 0.5 mg/ml, with minor differences in the distribution of nontrimeric species. rTNF-8 bound to purified 55and 75-kDa TNF receptors with high affinity. The binding of rTNF-8 to either receptor was analyzed on Scatchard plots and compared with that of rTNF-a.
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