Neun Barry W, Ilinskaya Anna N, Dobrovolskaia Marina A
Cancer Research Technology Program, Nanotechnology Characterization Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, P.O. Box B, Frederick, MD, 21702, USA.
Methods Mol Biol. 2018;1682:149-160. doi: 10.1007/978-1-4939-7352-1_13.
The complement system is a group of proteins, which function in plasma to assist the innate immunity in rapid clearance of pathogens. The complement system also contributes to coordination of the adaptive immune response. Complement Activation Related Pseudo Allergy or CARPA is a life-threatening condition commonly reported with certain types of drugs and nanotechnology-based combination products. While CARPA symptoms are similar to that of anaphylaxis, the mechanism behind this pathology does not involve IgE and is mediated by the complement system. In vitro assays using serum or plasma derived from healthy donor volunteers correlate with the in vivo complement-mediated reactions, and therefore are helpful in understanding the propensity of a given drug formulation to cause CARPA in patients. In the first edition of this book, we have described an in vitro method for qualitative assessment of the complement activation by nanomaterials using western blotting. Herein, we present a similar method utilizing enzyme-linked immunoassay for quantitative analysis of the complement activation, and we compare the performance of this approach to that of the qualitative western blotting technique. The revised chapter also includes new details about nanoparticle sample preparation.
补体系统是一组蛋白质，其在血浆中发挥作用，协助固有免疫快速清除病原体。补体系统也有助于协调适应性免疫反应。补体激活相关假过敏（CARPA）是一种常见于某些类型药物和基于纳米技术的组合产品的危及生命的病症。虽然CARPA症状与过敏反应相似，但这种病理背后的机制不涉及IgE，而是由补体系统介导。使用来自健康供体志愿者的血清或血浆进行的体外试验与体内补体介导的反应相关，因此有助于了解给定药物制剂在患者中引起CARPA的倾向。在本书的第一版中，我们描述了一种使用蛋白质印迹法对纳米材料补体激活进行定性评估的体外方法。在此，我们提出一种类似的方法，利用酶联免疫吸附测定法定量分析补体激活，并将该方法的性能与定性蛋白质印迹技术的性能进行比较。修订后的章节还包括有关纳米颗粒样品制备的新细节。
