The rabbit pyrogen test (RPT) is a safety test conducted as a part of mandatory requirements of regulatory agencies. RPT is currently performed for routine quality control (QC) by manufacturers and for national lot release of biological products, such as plasma-derived products. However, RPT involves the use of many rabbits, counter to the international efforts to minimize the use of animals in research. Furthermore, pyrogen amount cannot be discerned from the test results and the results may be considerably affected by various factors. Therefore, a need exists for substituting RPT with in vitro assays. As a viable alternative to RPT, we here established a rabbit monocyte activation test (RMAT) based on the human MAT in the European Pharmacopoeia. RMAT uses rabbit peripheral blood mononuclear cells as the source of monocytes instead of live animals. The test detected endotoxin, lipoteichoic acid, peptidoglycan, and zymosan with high sensitivity, showing high correlation with the in vivo RPT results. The results of RMAT and RPT testing of non-pyrogenic plasma-derived products were also consistent. Furthermore, RMAT showed satisfactory recovery rates in an interference test with product samples and spiked-in pyrogens. We conclude that RMAT could replace the existing RPT for routine QC.

兔热原测试 (RPT) 是作为监管机构强制性要求的一部分进行的安全测试。RPT 目前用于制造商的常规质量控制 (QC) 和生物产品的国家批次发布，例如血浆衍生产品。然而，RPT 涉及使用许多兔子，这与国际上尽量减少在研究中使用动物的努力背道而驰。此外，无法从测试结果中辨别热原量，并且结果可能受到各种因素的显着影响。因此，需要用体外替代 RPT化验。作为 RPT 的可行替代方案，我们在这里建立了基于欧洲药典中人类 MAT 的兔单核细胞活化试验 (RMAT)。RMAT使用兔外周血单核细胞代替活体动物作为单核细胞的来源。该测试对内毒素、脂磷壁酸、肽聚糖和酵母聚糖的检测灵敏度高，与体内RPT结果具有高度相关性。无热原血浆衍生产品的 RMAT 和 RPT 测试结果也一致。此外，RMAT 在对产品样品和掺入热原的干扰测试中显示出令人满意的回收率。我们得出结论，RMAT 可以取代现有的 RPT 用于常规 QC。