A pilot study comparing the purity, functionality and isoform composition of alpha-1-proteinase inhibitor (human) products
A pilot study comparing the purity, functionality and isoform composition of alpha-1-proteinase inhibitor (human) products
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DOI：
10.1185/030079905x46395
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发表时间：
2005-06-01
影响因子：
2.3
通讯作者：
Weeks, RL
中科院分区：
文献类型：
--
作者：
Cowden, DI;Fisher, GE;Weeks, RL
目的：α-1-蛋白酶缺乏症使受影响的个体易患早发性肺气肿，并且用来自合并的人血浆的α-1-蛋白酶抑制剂（A1-PI）治疗。本初步研究的目的是评估可能影响临床疗效、安全性和便利性的三种A1-PI产品（Aralast*、Prolastin（匕首）、Zemaira（双匕首））的分析参数。其中包括：制剂的纯度;杂质的性质;功能性;和同种型组成。使用反相和尺寸排阻色谱法高效液相色谱法（RP-HPLC和SEC-HPLC）、毛细管区带电泳（CZE）、十二烷基硫酸钠聚丙烯酰胺凝胶电泳、十二烷基硫酸钠毛细管凝胶电泳和Western印迹分析评价纯度。通过免疫比浊法确定蛋白质杂质的同一性;通过计算存在的活性A1-P1 mg（通过抗中性粒细胞弹性蛋白酶活性测定）与抗原A1-PI mg（通过免疫比浊法）的比值确定功能性;通过等电聚焦（IEF）确定A1-PI亚型模式的正态性。Zemaira的三个样品和Aralast和Prolastin各一个样品可用于分析。使用RP-HPLC分析，Zemaira平均纯度为99%，Aralast为70%，Prolastin小于62%。使用SEC-HPLC，Zemaira为95.98%单体，Prolastin为79.00%，Aralast为63.55%。Prolastin的活性/mg抗原A1-PI低于其他两种产品。CZE结果表明Aralast中异构体发生了变化，IEF结果证实了这一点。结论：Zemaira的纯度高于Aralast和Prolastin。Prolastin具有比Zemaira或Aralast更多的无活性A1-PI。在Aralast中，异构体比例似乎发生了改变。这项初步研究的结果值得进一步调查。
Objective: Alpha-1-proteinase deficiency predisposes affected individuals to early onset pulmonary emphysema, and is treated with an alpha-1-proteinase inhibitor (A1-PI) from pooled human plasma. The objective of this pilot study was to assess analytical parameters of the three A1-PI products (Aralast*, Prolastin(dagger), Zemaira(double dagger)) that may impact on clinical efficacy, safety, and convenience. These included: purity of the preparation; nature of impurities; functionality; and isoform composition.Methods: Purity was evaluated using reverse phase and size exclusion chromatography high performance liquid chromatography (RP-HPLC and SEC-HPLC), capillary zone electrophoresis (CZE), sodium dodecyl sulfate polyacrylamide gel electrophoresis, sodium dodecyl sulfate capillary gel electrophoresis and Western blot analysis. The identity of protein impurities was determined by immunonephelometry; functionality by calculating the ratio of mg active A1-P1 present (by anti-neutrophil elastase activity assay) to the mg antigenic A1-PI (by immunonephelometry); and normality of the A1-PI isoform pattern by isoelectric focusing (IEF). Three samples of Zemaira and one sample each of Aralast and Prolastin were available for analysis.Results: Zemaira had the highest specific activity. Using RP-HPLC analysis Zemaira averaged 99% purity, Aralast 70% and Prolastin less than 62%. Using SEC-HPLC Zemaira was 95.98% monomeric, Prolastin 79.00% and Aralast 63.55%. Prolastin had lower activity/mg antigenic A1-PI than the other two products. A shift in isoforms in Aralast was suggested by the results of CZE, and was confirmed by IEFConclusions: Zemaira demonstrated greater purity compared with Aralast and Prolastin. Prolastin had more inactive A1-PI than Zemaira or Aralast. Isoform ratios appeared to be altered in Aralast. The results from this pilot study warrant further investigation.