Purification of alpha 1 proteinase inhibitor from human plasma fraction IV-1 by ion exchange chromatography.
OBJECTIVE
Alpha-proteinase inhibitor (PI) protects the lungs from proteolytic damage caused by elastase and can be used to treat congenital emphysema. We describe an improved method of purification of alpha 1 PI from redissolved fraction IV-1 paste.


METHODS
The process used dimethylaminoethyl anion exchange chromatography, sulfopropyl cation exchange chromatography, virus inactivation by dry heat, and tri-n-butyl-phosphate/cholate treatment, followed by a second strong cation exchange chromatography. Optimizations of loading conditions for ion exchange chromatography at small scale (20-60 ml of suspension) are described. Virus inactivation was adjusted to provide the best yield of alpha 1 PI consistent with effective inactivation. The process has been effectively scaled up.


RESULTS
The final product was approximately 90% pure by SDS-PAGE, with a 60-70% yield from starting fraction IV-1 paste. The process has been characterized by methods including nonreduced SDS-PAGE, alpha 1 PI inhibition assay, and biuret protein assay.


CONCLUSIONS
The method described is an effective way of preparing large quantities of alpha 1 PI from fractionated plasma.
Chen
S X
SX
Technology, Bayer Corporation, Clayton, N.C., USA.
Hammond
D J
DJ
Lang
J M
JM
Lebing
W R
WR
D000838:Anions; D001798:Blood Proteins / Q000737:chemistry*; D002412:Cations; D020355:Cholates / Q000494:pharmacology; D002848:Chromatography, DEAE-Cellulose / Q000379:methods; D002852:Chromatography, Ion Exchange / Q000379:methods*; D003902:Detergents / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010755:Organophosphates / Q000494:pharmacology; D010865:Pilot Projects; D017434:Protein Structure, Tertiary; D012967:Sodium Dodecyl Sulfate; D012997:Solvents / Q000494:pharmacology; D013242:Sterilization / Q000379:methods; D013696:Temperature; D014775:Virus Activation; D014780:Viruses / Q000187:drug effects / Q000254:growth & development; D014867:Water; D000515:alpha 1-Antitrypsin / Q000737:chemistry / Q000302:isolation & purification*
D000838:Anions; D001798:Blood Proteins; D002412:Cations; D020355:Cholates; C032309:Cohn fraction IV; D003902:Detergents; D010755:Organophosphates; D012997:Solvents; D000515:alpha 1-Antitrypsin; D014867:Water; D012967:Sodium Dodecyl Sulfate; C009524:tributyl phosphate