Center for Biologics Evaluation and Research, US Food and Drug Administration, Silver Spring, MD, USA.
Nat Protoc. 2021 Aug;16(8):3981-4003. doi: 10.1038/s41596-021-00568-4. Epub 2021 Jul 2.
Thrombin generation (TG) assays are used widely to investigate both diseases and drugs that impact thrombosis and bleeding. TG assays were also instrumental in the identification of thrombogenic impurities in immune globulin products, which were associated with thrombotic adverse events in patients. TG assays are therefore now used by quality control laboratories of plasma derivative drug manufacturers and regulatory agencies responsible for the safety testing and release of immune globulin products. In this protocol, we describe a robust and sensitive version of the TG assay for quantitative measurement of thrombogenic activity in immune globulin products. Compared with the version of the assay commonly used in clinical laboratories that compares individual patient plasma samples with normal donor samples, our TG assay is suitable for quick (170-260 min) semiautomated analysis of multiple drug samples against the World Health Organization international standard for factor XIa. Commercially available reagents can be used for the assay, and it does not require specialized equipment. The protocol can be easily adapted for the measurement of the procoagulant activity of other biopharmaceuticals, e.g., coagulation factors.
凝血酶生成 (TG) 检测广泛用于研究影响血栓形成和出血的疾病和药物。TG 检测在鉴定免疫球蛋白产品中的血栓形成性杂质方面也发挥了重要作用，这些杂质与患者的血栓不良事件有关。因此，TG 检测现在被血浆衍生药物制造商的质量控制实验室和负责免疫球蛋白产品安全性测试和放行的监管机构使用。在本方案中，我们描述了一种用于定量测量免疫球蛋白产品中血栓形成活性的 TG 检测的稳健和灵敏版本。与在临床实验室中常用的版本相比，该版本的检测比较了个体患者的血浆样本与正常供体样本，我们的 TG 检测适合快速（170-260 分钟）半自动分析多个药物样本，以对抗世界卫生组织的因子 Xa 国际标准。可商购的试剂可用于该检测，并且不需要特殊设备。该方案可以轻松适应其他生物制药，例如凝血因子的促凝活性的测量。
