Research Laboratory, Nordland Hospital Bodø, Norway.
Mol Immunol. 2013 Dec 15;56(3):232-9. doi: 10.1016/j.molimm.2013.05.221. Epub 2013 Jun 17.
The importance of the complement system in clinical medicine has become evident during the last decades and complement therapeutics has now reached the clinic. Thus, there is an increased interest in and need for assays to evaluate complement activity and dysfunction. Pathologically increased complement activation can indirectly be evaluated by quantification of complement components, but in order to exactly measure such activation, assays for quantification of products formed during activation are required. Progress in this field is hampered by lack of standardization. Therefore, members of the International Complement Standardization Committee, a joint initiative of the International Complement Society and the International Union of Immunological Societies (IUIS), prepared a defined standard for application in assays for complement activation products. We here report on the production and properties of this International Complement Standard #2 (ICS#2). ICS#2 was made from a pool of sera from healthy blood donors (ICS#1) that was activated with a combination of heat-aggregated IgG and zymosan, and subsequently stabilized by adding EDTA and nafamostat mesylate. The protocol was optimized to make the standard applicable in the following activation product assays: C1rs-C1-inhibitor complexes, C4a, C4bc, C4d, Bb, C3bBbP, C3a, C3bc, C3dg, C5a and the soluble terminal C5b-9 complement complex (SC5b-9, TCC). ICS#2 was defined as containing 1000 complement activation units (CAU)/mL for all activation products measured. All activation products were stable after 10 times thawing and freezing and most of the activation products were stable during storage at 4°C for up to 21 days. ICS#2 was produced large-scale and is considered a valuable tool for standardization, calibration and reference control for complement activation assays, providing the necessary prerequisite for quality assessments between complement laboratories.
在过去的几十年中，补体系统在临床医学中的重要性已经显而易见，补体治疗现在已经进入临床。因此，人们对评估补体活性和功能障碍的检测方法的兴趣和需求日益增加。病理性补体激活程度可以通过补体成分的定量间接评估，但为了准确测量这种激活，需要定量检测激活过程中形成的产物。由于缺乏标准化，该领域的进展受到阻碍。因此，国际补体标准化委员会的成员（国际补体学会和国际免疫学会联合会的联合倡议）制定了一种用于补体激活产物检测的应用标准。我们在此报告这种国际补体标准品 #2（ICS#2）的生产和特性。ICS#2 由来自健康献血者血清的池（ICS#1）制成，该池与热聚集 IgG 和酵母聚糖联合激活，然后通过添加 EDTA 和萘莫司他甲磺酸盐稳定。该方案经过优化，可使标准品适用于以下激活产物检测：C1rs-C1 抑制剂复合物、C4a、C4bc、C4d、Bb、C3bBbP、C3a、C3bc、C3dg、C5a 和可溶性末端 C5b-9 补体复合物（SC5b-9、TCC）。ICS#2 被定义为所有测量的激活产物中含有 1000 个补体激活单位（CAU）/mL。所有激活产物在 10 次解冻和冷冻后均稳定，大多数激活产物在 4°C 下储存长达 21 天也稳定。ICS#2 是大规模生产的，被认为是补体激活检测标准化、校准和参考控制的有价值工具，为补体实验室之间的质量评估提供了必要的前提。
