摘要
作者分别用ConA-Sepharose 4B亲和层析法、DE-22 DEAE离子交换层析法、Blue-SepharoseCL-6B色素吸附层析法等方法初步纯化PHA-TPA协同刺激人外伤脾细胞产生的白细胞调节素(LR)及淋巴毒素(LT),并用DEAE-Sephadex A-50离子交换层析法进一步纯化经Con A-Sepharose 4B亲和层析法初步纯化的LR及LT。ConA-Sepharose 4B亲和层析法纯化LR及LT的回收率分别为28.1％及31.9％,最大提纯倍数分别为528.1倍及237.6倍;经DEAE-Sephadex A-50离子交换层析法进一步纯化,LR及LT的回收率分别为96％及92％,分别进一步提纯2.44倍及3.86倍;DF-22DEAE离子交换层析法LR及LT的回收率分别为87.5％及89.6％,最大提纯倍数分别为50.71倍:及78.88倍;Blue-Sepharose CL6B色素吸附法纯化LR及LT的回收率分别为53.75％及56.67％,最大提纯倍数分别为21.47倍及14.31倍,并研究了粗制及部分纯化LR或LT的稳定性。这些纯化方法的初步研究,为进一步连续高度纯化LR及LT创造了条件。
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.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
1989年第1期8-16,共9页
Academic Journal of Second Military Medical University
