High‐level production and isolation of human recombinant α1‐proteinase inhibitor in yeast

FEBS Letters

Bollen, A.²  Cabezón, T.¹  Harford, N.¹  Hoylaerts, M.²  Weyens, A.² 

[1] Biological Division, Smith Kline-RIT, rue de l'Institut 89, B-1330 Rixensart, Belgium;Génétique Appliquée, University of Brussels, rue de l'Industrie 24, B-1400 Nivelles, Belgium

关键词: α1-Proteinase;    Enzyme inhibitor;    α1-Antitrypsin isolation;    Elastase inhibitor substitution;    Emphysema;    (Yeast);    Expression vector;    Affinity chromatography;    α1-PI;    α1-proteinase inhibitor;    α1-antitrypsin;    r-α1-PI;    recombinant α1-PI;    PEG;    polyethylene glycol;    TDH3;    glyceraldehyde-3-phosphate dehydrogenase;    TNB;    4-nitrophenylsulfide 3;    3'-dicarbonic acid;   

DOI  :  10.1016/0014-5793(86)81391-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The cDNA coding for mature human α₁-proteinase inhibitor (α₁-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 α₁-PI. The production level is optimal when the recombinant plasmid carries the TDH promoter, the complete 2μ and the leu2D selection marker. Biologically active recombinant α₁-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-M _r, yeast material with polyethylene glycol 3300 followed by successive chromatography on DEAE-agarose, Zn-chelate agarose, κ-chain agarose, heparin-agarose and aminohexyl-agarose.

【 授权许可】

Unknown