Silicone Oil Microdroplets and Protein Aggregates in Repackaged Bevacizumab and Ranibizumab: Effects of Long-term Storage and Product Mishandling
Silicone Oil Microdroplets and Protein Aggregates in Repackaged Bevacizumab and Ranibizumab: Effects of Long-term Storage and Product Mishandling
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DOI：
10.1167/iovs.10-6431
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发表时间：
2011-02-01
影响因子：
4.4
通讯作者：
Carpenter, John F.
中科院分区：
文献类型：
--
作者：
Liu, Lu;Ammar, David A.;Carpenter, John F.
目的.为了定量从配制药房获得的重新包装的贝伐珠单抗以及在对照实验室实验中检测的贝伐珠单抗和雷珠单抗样品中的不溶性微粒和蛋白质聚集体的水平。重新包装的贝伐珠单抗购自4家外部复方药房。对于对照实验室研究，将贝伐珠单抗和安慰剂吸入塑料注射器中，并在-20 ° C、4 ° C和室温（有和无光照）下孵育12周。此外，使用含贝伐珠单抗的注射器模拟运输期间发生的机械冲击。通过颗粒表征技术定量颗粒计数和尺寸分布。采用分子排阻高效液相色谱法（SE-HPLC）测定贝伐珠单抗的单体和可溶性聚集体水平。来自配制药房的重新包装贝伐珠单抗的微粒计数范围很广（89，006 +/-56，406至602，062 +/-18，349/mL）。直接从原始玻璃小瓶中取样的贝伐珠单抗的微粒计数为63，839 +/- 349/mL。在重新包装的贝伐珠单抗中存在高达10%的单体损失。重新包装的贝伐珠单抗和安慰剂的实验室样本的初始颗粒计数分别为283，675 +/-60，494/mL和492，314 +/-389，361/mL。贝伐珠单抗和安慰剂样品的冻融导致> 120万个颗粒/mL。在所有重新包装的样品中，大部分微粒是由硅油引起的。SE-HPLC显示，与直接从小瓶中取出的贝伐珠单抗相比，在各种条件下在实验室中孵育的重新包装样品无显著差异。然而，反复冻融导致超过10%的单体损失。重新包装在塑料注射器中的贝伐珠单抗可能含有蛋白质聚集体，并被硅油微滴污染。冻融或其他处理不当会进一步增加颗粒污染物的水平。（Invest Ophthalmol维斯科学。2011; 52：1023-1034）DOI：10.1167/iovs.10-6431
PURPOSE. To quantify levels of subvisible particles and protein aggregates in repackaged bevacizumab obtained from compounding pharmacies, as well as in samples of bevacizumab and ranibizumab tested in controlled laboratory experiments.METHODS. Repackaged bevacizumab was purchased from four external compounding pharmacies. For controlled laboratory studies, bevacizumab and placebo were drawn into plastic syringes and incubated at -20 degrees C, 4 degrees C, and room temperature (with and without exposure to light) for 12 weeks. In addition, mechanical shock occurring during shipping was mimicked with syringes containing bevacizumab. Particle counts and size distributions were quantified by particle characterization technology. Levels of monomer and soluble aggregates of bevacizumab were determined with size-exclusion high-performance liquid chromatography (SE-HPLC).RESULTS. Repackaged bevacizumab from the compounding pharmacies had a wide range of particle counts (89,006 +/- 56,406 to 602,062 +/- 18,349/mL). Bevacizumab sampled directly from the original glass vial had particle counts of 63,839 +/- 349/mL. There was up to a 10% monomer loss in the repackaged bevacizumab. Laboratory samples of repackaged bevacizumab and placebo had initial particle counts, respectively, of 283,675 +/- 60,494/mL and 492,314 +/- 389,361/mL. Freeze-thawing of both bevacizumab and placebo samples led to > 1.2 million particles/mL. In all repackaged samples, most of the particles were due to silicone oil. SE-HPLC showed no significant differences for repackaged samples incubated in the laboratory under various conditions, compared with bevacizumab directly from vial. However, repeated freeze-thawing caused a more than 10% monomer loss.CONCLUSIONS. Bevacizumab repackaged in plastic syringes could contain protein aggregates and is contaminated by silicone oil microdroplets. Freeze-thawing or other mishandling can further increase levels of particle contaminants. (Invest Ophthalmol Vis Sci. 2011; 52: 1023-1034) DOI:10.1167/iovs.10-6431