Flow cytometry: a promising technique for the study of silicone oil-induced particulate formation in protein formulations.
Flow cytometry: a promising technique for the study of silicone oil-induced particulate formation in protein formulations.
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DOI：
10.1016/j.ab.2010.12.008
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发表时间：
2011-03-15
影响因子：
2.9
通讯作者：
Randolph TW
中科院分区：
文献类型：
--
作者：
Ludwig DB;Trotter JT;Gabrielson JP;Carpenter JF;Randolph TW
用于胃肠外给药的制剂中的不溶性微粒是生物制药行业关注的问题。然而，制剂组分（如用于预充式注射器润滑的硅油）可能会使不溶性微粒的监测和控制变得复杂，并且难以区分硅油微滴与聚集蛋白质形成的微粒。在本研究中，我们证明了流式细胞术能够分辨包含不溶性牛血清白蛋白（BSA）聚集体颗粒和吸附BSA的硅油乳液液滴的混合物。还利用流式细胞术研究硅油乳剂对含有表面活性剂、氯化钠或蔗糖的BSA、溶菌酶、阿巴西普或曲妥珠单抗制剂的稳定性的影响。为了帮助颗粒表征，通过用BODIPY® 493/503染色硅油和用Alexa Fluor® 647染色模型蛋白质来利用流式细胞术的荧光检测能力。流式细胞术分析显示，硅油乳剂通过蛋白质吸附到硅油液滴表面上诱导可溶性蛋白质的损失。添加表面活性剂可防止蛋白质吸附到硅油滴表面。尽管表面吸附曲妥珠单抗的油滴表现出絮凝，但由于暴露于硅油滴，形成的均匀蛋白质聚集体极少。本研究的结果证明了流式细胞术作为一种分析工具用于监测不溶性硅油滴对蛋白制剂稳定性的影响的实用性。
Subvisible particles in formulations intended for parenteral administration are of concern in the biopharmaceutical industry. However, monitoring and control of subvisible particulates can be complicated by formulation components, such as the silicone oil used for the lubrication of prefilled syringes, and it is difficult to differentiate microdroplets of silicone oil from particles formed by aggregated protein. In this study, we demonstrate the ability of flow cytometry to resolve mixtures comprising subvisible bovine serum albumin (BSA) aggregate particles and silicone oil emulsion droplets with adsorbed BSA. Flow cytometry was also utilized to investigate the effects of silicone oil emulsions on the stability BSA, lysozyme, abatacept or trastuzumab formulations containing surfactant, sodium chloride or sucrose. To aid in particle characterization, the fluorescence detection capabilities of Flow cytometry were exploited by staining silicone oil with BODIPY® 493/503 and model proteins with Alexa Fluor® 647. Flow cytometric analyses revealed that silicone oil emulsions induced the loss of soluble protein via protein adsorption onto the silicone oil droplet surface. Addition of surfactant prevented protein from adsorbing onto the surface of silicone oil droplets. There was minimal formation of homogeneous protein aggregates due to exposure to silicone oil droplets, although oil droplets with surface-adsorbed trastuzumab exhibited flocculation. The results of this study demonstrate the utility of flow cytometry as an analytical tool for monitoring the effects of subvisible silicone oil droplets on the stability of protein formulations.
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影响因子：
3.8
作者：
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