Fawaz Ibrahim, Schaz Simone, Boehrer Armin, Garidel Patrick, Blech Michaela
Boehringer Ingelheim Pharma GmbH & Co. KG, Innovation Unit, Pharmaceutical Development Biologicals, 88397 Biberach an der Riss, Germany.
Boehringer Ingelheim Pharma GmbH & Co. KG, Analytical Development Biologicals, CMC Statistics, 88397 Biberach an der Riss, Germany.
Eur J Pharm Biopharm. 2023 Apr;185:55-70. doi: 10.1016/j.ejpb.2023.01.017. Epub 2023 Jan 25.
Sub-visible particles (SVPs) in pharmaceutical products are a critical quality attribute, and therefore should be monitored during development. Although light obscuration (LO) and microscopic particle count tests are the primary pharmacopeial methods used to quantify SVPs, flow imaging methods like Micro-Flow Imaging (MFI™) appear to overcome shortcomings of LO such as limited sensitivity concerning smaller translucent SVPs in the size range < 10 µm. Nowadays, MFI™ is routinely utilized during development of biologicals. Oftentimes multiple devices are distributed across several laboratories and departments. This poses challenges in data interpretation and consistency as well as in the use of multiple devices for one purpose. In this study, we systematically evaluated seven MFI™ instruments concerning their counting and size precision and accuracy, using an inter-comparable approach to mimic daily working routine. Therefore, we investigated three different types of particles (i) NIST certified counting standards, (ii) protein-coated particles, and (iii) stress-induced particles from a monoclonal antibody. We compared the results to alternative particle detection methods: LO and Backgrounded Membrane Imaging (BMI). Our results showed that the precision and accuracy of particle count and size, as well as the comparability of instruments, depended on the particle source and its material properties. The various MFI™ instruments investigated showed high precision (<15 %) and data generated on different instruments were of the same order of magnitude within pharmacopeial relevant size ranges for NIST certified counting standards. However, we found limitations in the upper and lower detection limits, contrary to the limits claimed by the manufacturer. In addition, proteinaceous and protein-containing particles showed statistically significant differences in particle counts, while the measured particle diameters of all sizes were quite consistent.
药品中的亚可见颗粒（SVPs）是一项关键质量属性，因此在研发过程中应进行监测。尽管光阻法（LO）和显微镜颗粒计数测试是用于量化SVPs的主要药典方法，但诸如微流成像（MFI™）之类的流动成像方法似乎克服了光阻法的缺点，例如对于尺寸范围<10 µm的较小半透明SVPs，其灵敏度有限。如今，MFI™在生物制品研发过程中被常规使用。通常，多个设备分布在多个实验室和部门。这在数据解释和一致性以及将多个设备用于同一目的方面带来了挑战。在本研究中，我们采用一种可相互比较的方法来模拟日常工作流程，系统地评估了七台MFI™仪器在计数和尺寸精度及准确性方面的表现。因此，我们研究了三种不同类型的颗粒：（i）美国国家标准与技术研究院（NIST）认证的计数标准品，（ii）蛋白包被颗粒，以及（iii）单克隆抗体的应激诱导颗粒。我们将结果与其他颗粒检测方法进行了比较：光阻法和背景膜成像（BMI）。我们的结果表明，颗粒计数和尺寸的精度及准确性，以及仪器的可比性，取决于颗粒来源及其材料特性。所研究的各种MFI™仪器显示出高精度（<15%），并且在NIST认证计数标准品的药典相关尺寸范围内，不同仪器生成的数据在同一数量级。然而，与制造商声称的限值相反，我们发现了检测上限和下限的局限性。此外，蛋白质类和含蛋白质的颗粒在颗粒计数上显示出统计学上的显著差异，而所有尺寸的测量颗粒直径相当一致。
