Sx. Chen et al., PURIFICATION OF ALPHA(1) PROTEINASE-INHIBITOR FROM HUMAN PLASMA FRACTION IV-1 BY ION-EXCHANGE CHROMATOGRAPHY, Vox sanguinis, 74(4), 1998, pp. 232-241
Background and Objectives: alpha-proteinase inhibitor (PI) protects th
e lungs from proteolytic damage caused by elastase and can be used to
treat congenital emphysema. We describe an improved method of purifica
tion of alpha(1) PI from redissolved fraction IV-1 paste. Materials an
d Methods: The process used dimethylaminoethyl anion exchange chromato
graphy, 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 load
ing conditions for ion exchange chromatography at small scale (20-60 m
l of suspension) are described. Virus inactivation was adjusted to pro
vide the best yield of alpha(1) PI consistent with effective inactivat
ion. The process has been effectively scaled up. Results: The final pr
oduct was approximately 90% pure by SDS-PAGE, with a 60-70% yield from
starting fraction IV-1 paste. The process has been characterized by m
ethods including nonreduced SDS-PAGE, alpha(1) PI inhibition assay, an
d biuret protein assay. Conclusion: The method described is an effecti
ve way of preparing large quantities of alpha(1) PI from fractionated
plasma.