Chromatographic purification of human alpha 1 proteinase inhibitor from dissolved Cohn fraction IV-1 paste.
A novel chromatographic process for purification of alpha 1 proteinase inhibitor (alpha 1-PI) from Cohn fraction IV-1 paste is described. This process has been successfully scaled up to 50-1 columns. It involves DEAE chromatography, sulfopropyl (S) cation chromatography, tri-n-butyl phosphate (TNBP)-cholate treatment, a second S cation chromatography, freeze-drying and dry-heat. The process has been optimized for purity, yield, lipid removal, chemical usage and water consumption. Filtration after TNBP-cholate treatment plays a key role in ensuring a low lipid content in the final product. Pre-equilibration with high salt buffer is necessary to reduce the water consumption significantly during the ion-exchange chromatography equilibration step. The final product is approximately 95% pure by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with a 64% to 70% yield from IV-1 paste.
Chen
S X
SX
Bayer Corporation, Raleigh, NC 27606, USA.
Hammond
D J
DJ
Klos
A M
AM
Wood
W D
WD
Wydick
J E
JE
Lebing
W R
WR
D001798:Blood Proteins / Q000737:chemistry*; D002784:Cholesterol / Q000032:analysis / Q000302:isolation & purification; D019826:Cholic Acid; D002793:Cholic Acids / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D002848:Chromatography, DEAE-Cellulose / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D010755:Organophosphates / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D015203:Reproducibility of Results; D015842:Serine Proteinase Inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D000515:alpha 1-Antitrypsin / Q000302:isolation & purification* / Q000378:metabolism
D001798:Blood Proteins; D002793:Cholic Acids; C032309:Cohn fraction IV; D010755:Organophosphates; D015842:Serine Proteinase Inhibitors; D000515:alpha 1-Antitrypsin; C009524:tributyl phosphate; D002784:Cholesterol; D019826:Cholic Acid