Analytical Research and Development, Ono Pharmaceutical Co., Ltd., 3-1-1 Sakurai Shimamoto-cho, Mishima-gun, Osaka, 618-8585, Japan; Graduate School of Science, Technology and Innovation & Engineering Biology Research Center, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, 657-8501, Japan.
Graduate School of Science, Technology and Innovation & Engineering Biology Research Center, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe, 657-8501, Japan.
Biologicals. 2020 Jul;66:1-8. doi: 10.1016/j.biologicals.2020.06.006. Epub 2020 Jul 10.
Bioburden control in the manufacturing facility is a serious concern regarding biologics due to the possibility of an significant economic impact due to batch failure from a bioburden incident. As a case study on effectively establishing a microbiological environmental monitoring program for cleanrooms, we focused on Time-lapse Shadow Image Analysis as a kind of Rapid Microbiological Method. In this study, the superior rapidity and accuracy were indicated for reference strains and environmental microbial on both 90 mm plate and RODAC plate at 25-30 °C. Especially superior performance in the counting was observed for B.subtilis, P.aeruginosa and A.brasiliensis. The first and the median of colony detection speed for environmental microbial were 12 h and 26 h, respectively. The colony detection rate was 90% at 40 h incubation. Additionally, the characterization of swarming behavior was recognized based on time-lapse image acquisition data at 30 min intervals. This case study indicated that the application for environmental monitoring can contribute to reducing the bioburden excursion risk due to both the rapid detection of colonies and real-time detection for swarming behavior. TSIA would be more acceptable and easier option for biologics due to providing simple interpretations for the results and reducing the time consumption.
由于生物负荷事件可能导致批次失败，从而对生物制品产生重大的经济影响，因此，在生产设施中控制生物负荷是一个严重的问题。作为一个关于如何有效建立洁净室微生物环境监测计划的案例研究，我们专注于时间 lapse 阴影图像分析（Time-lapse Shadow Image Analysis）作为一种快速微生物方法。在这项研究中，在 25-30°C 下，90mm 平板和 RODAC 平板上的参考菌株和环境微生物均显示出快速性和准确性优势。尤其是枯草芽孢杆菌（B.subtilis）、铜绿假单胞菌（P.aeruginosa）和巴西曲霉（A.brasiliensis）的计数性能更优。环境微生物的菌落检测速度的第一和中位数分别为 12 小时和 26 小时。40 小时孵育时，菌落检测率为 90%。此外，还可以基于 30 分钟间隔的时间 lapse 图像采集数据来识别迁徙行为的特征。这项案例研究表明，由于能够快速检测菌落并实时检测迁徙行为，该方法在环境监测中的应用有助于降低生物负荷偏移风险。由于能够提供简单的结果解释并减少时间消耗，因此时间 lapse 阴影图像分析（TSIA）对于生物制品来说是更可接受和更容易的选择。
