Center for Pharmaceuticals Research and Development, Havana, Cuba.
ALTEX. 2011;28(3):227-35. doi: 10.14573/altex.2011.3.227.
Disadvantages of the regulatory pyrogen test to assure safety of the end-product Human Serum Albumin (HSA) for parenteral use call for the implementation of an alternative test. In the current study, 16 HSA batches were assayed for pyrogens in parallel with the Rabbit Pyrogen Test, conventional and endotoxin-specific LAL assay and monocyte activation test (MAT). It was found that all HSA batches were contaminated with (1,3)-beta-glucans, which interfere with the conventional LAL. Endotoxin-specific LAL was not suitable to test HSA due to unacceptable endotoxin recovery. Experiments combining polymyxin B and MAT demonstrated that pyrogenic batches were mainly contaminated with endotoxins. However, endotoxin-specific LAL failed to detect one of them. The contaminating (1,3)-beta-glucans enhanced the MAT/IL-6 response to endotoxin, but not that of MAT/IL-1beta. The endotoxin equivalent concentrations obtained using the IL-6 readout were usually higher than those using IL-1beta, probably owing to the direct induction of IL-6 release from monocytes by (1,3)-beta-glucans. The MAT correlates with the rabbit pyrogen test, providing a higher safety level for pyrogenicity testing of HSA and probably other therapeutic proteins.
用于保证人血清白蛋白（HSA）用于注射用药的安全性的监管热原检测的缺点，要求实施替代检测。在本研究中，16 批 HSA 与兔热原检测、常规和内毒素特异性 LAL 检测以及单核细胞激活检测（MAT）平行进行热原检测。结果发现，所有 HSA 批次均被（1,3）-β-葡聚糖污染，这会干扰常规 LAL。由于内毒素回收率不可接受，内毒素特异性 LAL 不适合用于检测 HSA。结合多粘菌素 B 和 MAT 的实验表明，致热批次主要被内毒素污染。然而，内毒素特异性 LAL 未能检测到其中的一个批次。污染的（1,3）-β-葡聚糖增强了 MAT/IL-6 对内毒素的反应，但不能增强 MAT/IL-1β 的反应。使用 IL-6 读数获得的内毒素等效浓度通常高于使用 IL-1β 获得的浓度，这可能是由于（1,3）-β-葡聚糖直接诱导单核细胞释放 IL-6。MAT 与兔热原检测相关，为 HSA 及可能的其他治疗性蛋白的热原检测提供了更高的安全性水平。
