人血浆Cohn法组分Ⅳ沉淀的综合利用研究

Comprehensive utilization research of Cohn fraction Ⅳ precipitate of human plasma

目的采用全层析工艺从废弃的Cohn法组分Ⅳ沉淀(Cohn fraction Ⅳ precipitate, Cohn F Ⅳ)中提取抗凝血酶Ⅲ(antithrombin Ⅲ, AT Ⅲ)、α1-抗胰蛋白酶(α1-antitrypsin,α1-AT)和人血白蛋白进行综合利用。方法将Cohn F Ⅳ溶解并澄清处理后进行SD法病毒灭活,得到上柱前料液,再依次采用阴离子交换层析、肝素亲和层析+α1-AT Select串联亲和层析、疏水层析以及分子筛层析后分别得到AT Ⅲ和α1-AT的纯品以及主要含有人血白蛋白的组分。对AT Ⅲ和α1-AT分别进行纯度、活性以及肝素亲和力等方面的检测,并评价其质量属性。结果采用此方法获得的AT Ⅲ纯度可达99%以上,比活达10 IU/mg以上,肝素亲和力达80%以上;α1-AT纯度同样可达99%以上,比活达1.5 PU/mg以上。结论本研究所述方法可成功将Cohn F Ⅳ中的AT Ⅲ、α1-AT以及人血白蛋白等几种目前最具备提取利用价值的蛋白进行有效的分离,在人血浆综合利用方面具有较高的利用价值。

Objective Using all-chromatography technology to comprehensively extract and utilize antithrombin Ⅲ(AT Ⅲ), α1-antitrypsin(α1-AT) and albumin from discarded Cohn fraction Ⅳ precipitate. Methods Dissolving and clarifying the Cohn fraction Ⅳ precipitate and then inactivating virus by SD method to obtain the pre-column feed solution.In turn, using anion exchange chromatography, heparin+α1-AT Select tandem affinity chromatography, hydrophobic interaction chromatography and size exclusion chromatography to obtain purified AT Ⅲ, α1-AT and component of albumin. AT Ⅲ and α1-AT were examined for purity, activity and heparin affinity to evaluate its quality attributes. Results Using our method to obtain pure AT Ⅲ product, which purity was over 99% and specific activity was up to 10 IU/mg or more. The heparin-binding fraction was over 80%, similarly, the purity of α1-AT was up to 99% and the specific activity was up to 1.5 PU/mg or more. Comparing with similar products in currently foreign market, our products had higher quality standards. Conclusion Our method in this study could successfully separate AT Ⅲ, α1-AT and albumin from fraction Ⅳ precipitate, these proteins were thought to be the most extracted and use of valuable protein. Moreover, our products could achieve higher quality standards, therefore, which had a high value in comprehensive utilization of human plasma.