α_1-抗胰蛋白酶制剂的制备及其病毒灭活  

Preparation and Virus Inactivation of α_1 Antitrypsin

作　　者：朱威[1] 祝慈芳[1] 刁武萍[1] 周继东[1] 吴玮[1] 

机构地区：[1]上海生物制品研究所,上海200052

出　　处：《中国生物制品学杂志》2001年第2期97-101,共5页Chinese Journal of Biologicals

摘　　要：目的 从Ｃｏｈｎ　ＦⅣ中提纯α１－抗胰蛋白酶（α１－Ａｎｔｉｔｒｙｐｓｉｎ　α１－ＡＴ，制备较高纯度α１－ＡＴ制剂。方法 对Ｃｏｈｎ ＦⅣ抽提液经沉淀、超滤、离子交换及凝胶过滤，提纯 α１－ＡＴ用巴氏法（液态，６０℃，１０ｈ加热）作病毒灭活， 并考察其效果。用免疫单扩散、双向交叉免疫电泳、ＳＤＳ－ＰＡＧＥ及合成基质法检测α１－ＡＴ蛋白及生物活性。用区带 扫描法测定α１－ＡＴ制剂的纯度。结果３批试制的α１－ＡＴ制剂的纯度为９１．３±１．５２％，比活０．４９±０．０８μ／ｍｇ，比活性 比抽提液提高了２７．４倍。巴氏法处理可使ＶＳＶ、Ｓｉｎｄｂｉｓ、Ｐｏｌｉｏ　Ｉ型疫苗病毒分别降低１０．０±０．３，１０．０±０．３１及７．１１ ±０．３ｌｇＴＣＩＤ５０／ｍｌ，但仍能检出Ｐｏｌｉｏ型疫苗病毒。结论 可从 Ｃｏｈｎ ＦⅣ中制得较高纯度的 α１－ＡＴ制剂，巴氏法能对 α１－ＡＴ制剂作有效的病毒灭活处理。Objective To purify α1 antitrypsin(α１ -AT )from Calm FIV and observe the efficiency of virus inactivation in it. Methods α１ -AT was purified from Cohn FIV by precipitation ,ultrafiltration, ion-exchange and gel filtration, and the virus in it was inactivated by pasteurization. The specific activity of α１ -AT was detected by single immumodiffusion , cress immunoelectrophoresis, SDS-PAGE and peptide chromogenic substrates method, and the purity of it by band scanning method.Results The purity of three lots of α１-AT was(91.3± 1.52)%,and the specific activity of it was 0.49 ± 0.08u/mg,which was 27.4 times higher than that of source material before purification.The titers of model viruses VSV,Sindbis,poliovirus type I decreased by 10.0 ± 0.3,10.0± 0.3 and 7.11±0.31gTCID50/ml respectively,while poliovirus type I still could be detected out after virus inactivation. Conclusion The purification procedure can be used as an effective way to prepare highly purified α１ -AT, and the viruses in α１ -AT can be effectively inactivated by pasteurization.

关 键 词：Α1-抗胰蛋白酶 病毒灭活 CohnFⅣ 

分 类 号：TQ464.8[化学工程—制药化工] R977.3[医药卫生—药品]