Renaturation and purification of recombinant tissue-type plasminogen activator expressed in E. coli.
Under the control of trp promoter, human tissue-type plasminogen activator was expressed in E. coli in the form of inclusion body. The recombinant t-PA was recovered for renaturation from preparative native PAGE, gel by zinc acetate staining and electroelution. After renaturation in vitro, the recombinant t-PA was purified by benzamidine affinity chromatography and lysine affinity chromatography. The purified t-PA showed homogeneous on silver-stained SDS-PAGE gel, with a specific activity of 240,000 I.U./mg protein.
Hua
Z C
ZC
Department of Biochemistry, Nanjing University, People's Republic of China.
D002846:Chromatography, Affinity / Q000379:methods; D004926:Escherichia coli / Q000235:genetics; D005337:Fibrin / Q000378:metabolism; D006801:Humans; D002479:Inclusion Bodies; D011489:Protein Denaturation; D017510:Protein Folding; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D010959:Tissue Plasminogen Activator / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D005337:Fibrin; D010959:Tissue Plasminogen Activator