PARTIAL PURIFICATION OF LEUKOREGULIN AND LYMPHOTOXIN PRODUCED BY PHYTOHEMAGGLUTiNIN AND DITERPENE ESTER ACTIVATED HUMAN SPLEEN CELLS
- VernacularTitle:人白细胞调节素与淋巴毒素纯化方法的研究
- Author:
Jian NI
- Publication Type:Journal Article
- Keywords:
leukoregulin.;
lymphotoxin;
purification
- From:
Academic Journal of Second Military Medical University
1982;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
In order to provide highly purified preparation of human ieukoregulin (LR) for the exploration of its anticancer mschatiism, crude preparations of LR and lymphotoxin(LT) derived from human spleen cell cultures induced with phytohem-agglutinin and diterp;ne ester l2-O-tetradecanoylphorbol-i3-acetate were purified by using four purification nmhois : (a) Con A-Sepharose 48 affinity chromatography, by which crude human LR or LT was purified to maximum specific activity of 85 333 U/mg protein or 13333 U/mg protein with a recovery of 28.1% or 31.9%, respectively 5 (b) DEAE-Sjphadex A-50 ion-exchange chromatography, by which partially purified LR or LT obtained after Con A-Sepharose 4B affinity chromatography was further purified 2.44-fold or 3.8o-fold with a recovery of 96% or 92%, respectively ; (c) DEAE-22-celluiose ion-exchange chromatography, by which crude human LR or LT was purified to maximum specific activity of 11 294 U/mg protein or 13 176 U/mg protein with a recovery of 87.5% or 89.6%.respectively ; (d) Blue-Sepharose CL-6B affinity chromatography, by which crude human LR or LT was purified to maximum specific activity of 51.89 U/mg protein or 2594 U/mg protein with a recovery of 53.75% or 5t3.G7%, respectively.Our data indicate that Con A-Sepharose 4B affinity chromatography, DEAE-Sephadex A-50 ion-exchange chromatography.DEAE-22-cellulose ion-exchange chromatography and Blue-Sepharose CL-6B affinity chromatography may be useful in the purification of LR or LT.
