Characterization and quantitation of aggregates and particles in interferon- products: Potential links between product quality attributes and immunogenicity
Characterization and quantitation of aggregates and particles in interferon- products: Potential links between product quality attributes and immunogenicity
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DOI：
10.1002/jps.23415
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发表时间：
2013-03-01
影响因子：
3.8
通讯作者：
Carpenter, John F.
中科院分区：
文献类型：
--
作者：
Barnard, James G.;Babcock, Ken;Carpenter, John F.
干扰素- (IFN-)产品已用于多发性硬化症的治疗多年，包括重组IFN- 1b （Betaseron (R)）和IFN- 1a (Avonex （R)和Rebif (R)）。所有这三种产品都会导致中和抗体（nab）的形成，并导致患者疗效的丧失，但程度不同。在几项临床试验中，报告的中和抗体形成率为22% - 47% (Betaseron (R)), 5% - 35% （Rebif (R)）和2% - 13% （Avonex (R)）。在目前的研究中，所有的产品都是从药店购买的，聚集体使用粒径排除层析（SEC）、分析性超离心、凝胶电泳和点印迹免疫测定进行表征和/或定量。通过微流成像、颗粒跟踪分析和共振质量测量进行颗粒表征和计数。用人血清白蛋白配制的Betaseron (R)和Rebif (R)聚集的蛋白质和颗粒数量最多（例如，SEC显示9% - 15%的高分子量物种，血流成像显示bbb10万颗粒/mL）。结果表明，Avonex (R)的聚集蛋白含量最少，单体含量为0.95%，且Avonex (R)中检测到的颗粒主要为硅油滴。这些结果强烈表明，蛋白质聚集体和颗粒含量是关键的产品质量属性，在一个给定的产品的倾向，引发生产的nab在患者。(3)中国医药科学，2013 (2):1 - 4
Interferon- (IFN-) products have been used for many years in the treatment of multiple sclerosis and include recombinant IFN--1b (Betaseron (R)) and IFN--1a (Avonex (R) and Rebif (R)). All three products lead to the formation of neutralizing antibodies (NAbs) and resulting loss of efficacy in patients but to different extents. Across several clinical trials, the reported rates of neutralizing-antibody formation were 22%47% (Betaseron (R)), 5%35% (Rebif (R)), and 2%13% (Avonex (R)). In the current study, all products were purchased from the pharmacy and aggregates were characterized and/or quantified using size-exclusion chromatography (SEC), analytical ultracentrifugation, gel electrophoresis, and dot-blotting immunoassays. Particle characterization and counting were performed using microflow imaging, particle tracking analysis, and resonant mass measurement. Betaseron (R) and Rebif (R), which are formulated with human serum albumin, had the greatest amount of aggregated protein and particles (e.g., 9%15% high molecular weight species by SEC and >100,000 particles/mL by flow imaging). Avonex (R) was found to have the least amount of aggregated protein, with >95% monomer content by both SEC and analytical ultracentrifugation, and the particles detected in Avonex (R) were determined to be primarily silicone oil droplets. These results strongly suggest that protein aggregate and particle contents are key product quality attributes in a given product's propensity to elicit the production of NAbs in patients. (c) 2012 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 102:915928, 2013