High-level production and isolation of human recombinant alpha 1-proteinase inhibitor in yeast.
The cDNA coding for mature human alpha 1-proteinase inhibitor (alpha 1-PI) has been inserted into a variety of yeast expression vectors. Yeast cells transformed with these plasmids were then assayed for the production of mature, unglycosylated alpha 1-PI. The production level is optimal when the recombinant plasmid carries the TDH promoter, the complete 2mu and the leu2D selection marker. Biologically active recombinant alpha 1-PI can be purified either analytically, by affinity chromatography using a monoclonal antibody, or on a large scale, by a procedure involving precipitation of high-Mr yeast material with polyethylene glycol 3300 followed by successive chromatography on DEAE-agarose, Zn-chelate agarose, kappa-chain agarose, heparin-agarose and aminohexyl-agarose.
D001798:Blood Proteins / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation; D006801:Humans; D010957:Plasmids; D011994:Recombinant Proteins / Q000302:isolation & purification*; D015003:Yeasts; D000515:alpha 1-Antitrypsin
D001798:Blood Proteins; D011994:Recombinant Proteins; D000515:alpha 1-Antitrypsin