Limentani S A, Gowell K P, Deitcher S R
Department of Medicine, New England Medical Center Hospitals, Boston, MA 02111, USA.
Thromb Haemost. 1995 Apr;73(4):584-91.
This study employed sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis and immunoblotting to assess the purity of seven high purity factor IX concentrates: Aimafix (Aima), AlphaNine-SD (Alpha Therapeutic), Factor IX VHP (Biotransfusion), Immunine (Immuno), Mononine (Armour Pharmaceutical), Nanotiv (Kabi Pharmacia), and 9MC (Blood Products Laboratory). The mean specific activity of these products ranged from 68 U factor IX/mg (Aimafix) to 246 U factor IX/mg (Mononine). SDS-PAGE analysis showed that the highest purity product, Mononine, had a single contaminating band under non-reducing conditions. Two additional bands were detected when this product was analyzed under reducing conditions. All other products had multiple contaminating bands that were more apparent under reducing than non-reducing conditions. The immunoblot for factor IX showed a dominant factor IX band for all products. In addition, visible light chain of factor IX was detected for AlphaNine-SD, Factor IX VHP, Immunine, Mononine, Nanotiv, and 9MC, suggesting that the factor IX in these products had undergone partial activation to factor IXa. Another contaminating band was visible at 49,500 for all of the products except 9MC. In addition to this band, high molecular weight contaminants were apparent for some products, most notably AlphaNine-SD. The identity of these bands is unknown. Immunoblotting failed to demonstrate factor VII as a contaminant of any of the high purity products, although factor VIIa could be detected in some lots of Immunine, Nanotiv, and 9MC by a clot-based assay. Factor X contaminated Aimafix, AlphaNine-SD, Factor IX VHP, Immunine, Nanotiv, and 9MC, but activation products of factor X were not detected.(ABSTRACT TRUNCATED AT 250 WORDS)
本研究采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析和免疫印迹法，评估了七种高纯度凝血因子IX浓缩剂的纯度：Aimafix（爱脉隆）、AlphaNine-SD（阿尔法治疗公司）、Factor IX VHP（生物输血公司）、Immunine（免疫公司）、Mononine（阿莫尔制药公司）、Nanotiv（卡比 Pharmacia 公司）和9MC（血液制品实验室）。这些产品的平均比活性范围为68 U凝血因子IX/毫克（Aimafix）至246 U凝血因子IX/毫克（Mononine）。SDS-PAGE分析表明，纯度最高的产品Mononine在非还原条件下有一条单一的污染带。在还原条件下分析该产品时，检测到另外两条带。所有其他产品都有多个污染带，在还原条件下比非还原条件下更明显。凝血因子IX的免疫印迹显示所有产品都有一条主要的凝血因子IX带。此外，在AlphaNine-SD、Factor IX VHP、Immunine、Mononine、Nanotiv和9MC中检测到了可见的凝血因子IX轻链，表明这些产品中的凝血因子IX已部分激活为凝血因子IXa。除9MC外，所有产品在49,500处都可见另一条污染带。除了这条带外，一些产品还明显存在高分子量污染物，最显著的是AlphaNine-SD。这些带的身份未知。免疫印迹未能证明凝血因子VII是任何高纯度产品的污染物，尽管通过基于凝血的检测方法在一些Immunine、Nanotiv和9MC批次中可检测到凝血因子VIIa。凝血因子X污染了Aimafix、AlphaNine-SD、Factor IX VHP、Immunine、Nanotiv和9MC，但未检测到凝血因子X的激活产物。（摘要截断于250字）
