Updated Method for In Vitro Analysis of Nanoparticle Hemolytic Properties
Updated Method for In Vitro Analysis of Nanoparticle Hemolytic Properties
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DOI：
10.1007/978-1-4939-7352-1_9
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发表时间：
2018-01-01
期刊：
影响因子：
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通讯作者：
Dobrovolskaia, Marina A.
中科院分区：
文献类型：
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作者：
Neun, Barry W.;Ilinskaya, Anna N.;Dobrovolskaia, Marina A.
溶血是红细胞 (RBC) 受损，导致含铁血红蛋白释放到血浆中。先前开发并描述了一种体外测定法，用于分析纳米颗粒溶血特性。在这里，我们提出了原始协议的修订版本。在此方案中，分析物纳米粒子和对照物在血液中孵育。通过离心去除未受损的红细胞，受损红细胞释放的血红蛋白通过与 Drabkin 试剂一起孵育转化为氰化高铁血红蛋白。通过分光光度法测量上清液中氰化高铁血红蛋白的量。将测得的吸光度与标准曲线进行比较，以确定上清液中血红蛋白的浓度。然后将测得的血红蛋白浓度与总血红蛋白浓度进行比较，以获得纳米颗粒诱导的溶血的百分比。该修订版包括有关纳米颗粒样品制备、体外研究纳米颗粒浓度选择的更新详细信息、有关测定控制的更新详细信息以及有关纳米颗粒干扰体外溶血测定的案例研究。
Hemolysis is damage to red blood cells (RBCs), which results in the release of the iron-containing protein hemoglobin into plasma. An in vitro assay was developed and described earlier for the analysis of nanoparticle hemolytic properties. Herein, we present a revised version of the original protocol. In this protocol, analyte nanoparticles and controls are incubated in blood. Undamaged RBCs are removed by centrifugation and hemoglobin, released by the damaged erythrocytes, is converted to cyanmethemoglobin by incubation with Drabkin's reagent. The amount of cyanmethemoglobin in the supernatant is measured by spectrophotometry. This measured absorbance is compared to a standard curve to determine the concentration of hemoglobin in the supernatant. The measured hemoglobin concentration is then compared to the total hemoglobin concentration to obtain the percentage of nanoparticle-induced hemolysis. The revision includes updated details about nanoparticle sample preparation, selection of nanoparticle concentration for the in vitro study, updated details about assay controls and case studies about nanoparticle interference with the in vitro hemolysis assay.