High-level production and isolation of human recombinant α1-proteinase inhibitor in yeast
| Autor: | A. Weyens, Teresa Cabezón, N. Harford, Andrew W. Bollen, Marc Hoylaerts |
|---|---|
| Jazyk: | angličtina |
| Předmět: |
α1-Proteinase
α1-Antitrypsin isolation Expression vector medicine.drug_class Biophysics Biology Monoclonal antibody Affinity chromatography Biochemistry law.invention chemistry.chemical_compound Plasmid Structural Biology law Complementary DNA Yeasts Genetics medicine Humans Molecular Biology Emphysema Cell Biology Blood Proteins Enzyme inhibitor Molecular biology Yeast Recombinant Proteins chemistry Gene Expression Regulation Elastase inhibitor substitution alpha 1-Antitrypsin (Yeast) Recombinant DNA Agarose Electrophoresis Polyacrylamide Gel Plasmids |
| Zdroj: | FEBS Letters. (1):83-87 |
| ISSN: | 0014-5793 |
| DOI: | 10.1016/0014-5793(86)81391-6 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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