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曲拉通X-100对溶剂/去污剂处理血浆中α2-抗纤溶酶(SERPINF2)活性的影响。

Impact of Triton X-100 on alpha 2-antiplasmin (SERPINF2) activity in solvent/detergent-treated plasma.

作者信息

Burnouf Thierry, Goubran Hadi Alphonse, Radosevich Miryana, Sayed Makram A, Gorgy George, El-Ekiaby Magdy

机构信息

Research Department, Human Plasma Product Services, 18 rue Saint Jacques, 59800 Lille, France.

出版信息

Biologicals. 2007 Oct;35(4):349-53. doi: 10.1016/j.biologicals.2007.03.002. Epub 2007 Jul 26.

Abstract

Large-pool solvent/detergent (SD) plasma for transfusion exhibits reduced alpha 2-antiplasmin (alpha2-AP; SERPINF2) functional activity. The reason for the loss of alpha2-AP has not been described and could be due to the SD incubation itself and/or to the processing steps implemented to remove the solvent and the detergent. We have studied alpha2-AP activity during six down-scale preparations of plasma virally-inactivated by 1% (v/v) TnBP combined with two different non-ionic detergents, either 1% Triton X-100 or 1% Triton X-45, at 31 degrees C for 4h. The SD-treated plasmas were then extracted with 7.5% (v/v) soybean oil, centrifuged at 3800 x g for 30 min, and subjected to hydrophobic interaction chromatography (HIC) to remove the SD agents. Control runs without TnBP and Triton were performed to evidence possible impacts of each process step on alpha2-AP activity. TnBP, Triton X-100, and Triton X-45 were measured at all stages of the processes to evaluate potential interferences with the alpha2-AP assay. Alpha 2-AP activity was about 10% that of starting plasma after 1% TnBP-1% Triton X-100 incubation and about 50% after oil extractions, centrifugation, and HIC. By contrast about 73% of the antiplasmin activity was found after the incubation with 1% TnBP and 1% Triton X-45, 88% after removal of the SD agents by oil extractions, 90% after centrifugation and 92% after HIC. The control runs performed without SD agents showed that the process steps did not affect the alpha2-AP activity. In conclusion, the agent altering alpha2-AP activity in SD-plasma is Triton X-100. The choice of detergents for the SD viral inactivation of therapeutic plasma fractions used in patients at risk of fibrinolysis should consider the impact on alpha2-AP activity.

摘要

用于输血的大池溶剂/去污剂(SD)血浆显示α2-抗纤溶酶(α2-AP;丝氨酸蛋白酶抑制剂F2)功能活性降低。α2-AP活性丧失的原因尚未阐明,可能是由于SD孵育本身和/或为去除溶剂和去污剂而实施的处理步骤所致。我们研究了在6次小规模制备过程中α2-AP的活性,这些血浆在31℃下用1%(v/v)TnBP与两种不同的非离子去污剂(1% Triton X-100或1% Triton X-45)联合处理4小时进行病毒灭活。然后用7.5%(v/v)大豆油提取经SD处理的血浆,以3800×g离心30分钟,并进行疏水相互作用色谱(HIC)以去除SD剂。进行了不添加TnBP和Triton的对照实验,以证明每个工艺步骤对α2-AP活性的可能影响。在工艺的所有阶段都对TnBP、Triton X-100和Triton X-45进行了测量,以评估对α2-AP测定的潜在干扰。在1% TnBP - 1% Triton X-100孵育后,α2-AP活性约为起始血浆的10%,在油提取、离心和HIC后约为50%。相比之下,在与1% TnBP和1% Triton X-45孵育后,约73%的抗纤溶酶活性被保留,在通过油提取去除SD剂后为88%,离心后为90%,HIC后为92%。不使用SD剂进行的对照实验表明,工艺步骤不会影响α2-AP活性。总之,在SD血浆中改变α2-AP活性的试剂是Triton X-100。对于有纤溶风险的患者使用的治疗性血浆组分进行SD病毒灭活时,去污剂的选择应考虑对α2-AP活性的影响。

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