Spaeth Scott, Tran Quynh, Liu Zhiyun
Bristol-Myers Squibb, Biologics Lead Discovery, Redwood City, CA, USA.
Bristol-Myers Squibb, Biologics Lead Discovery, Redwood City, CA, USA
PDA J Pharm Sci Technol. 2018 Nov-Dec;72(6):574-583. doi: 10.5731/pdajpst.2018.008557. Epub 2018 Jun 27.
This study compared an adenosine triphosphate (ATP)-based bioluminescence rapid microbial method (RMM) with a conventional sterility method for biologics sample testing. The RMM is based on a comparison of ATP levels in inoculated and uninoculated microbiological growth medium samples following growth enrichment incubation. The biologics samples qualified in this study were recombinant monoclonal antibodies and hybridoma cell culture supernatants. Initially, the lot-to-lot variation in background ATP of these samples posed significant challenges. Two strategies to increase the signal-to-noise ratio (positive result/background ATP) were evaluated: enzyme-based signal amplification and reduction of the broth-based noise through broth selection. Following qualification of the RMM for antibody and cell culture samples, the RMM was also utilized for rapid screening of several sources of purified water. This ATP-based RMM has proved invaluable in routine testing of diverse biologics samples at our discovery research site and plays a key role in the investigation of contaminated samples. Biologics research laboratories routinely conduct sterility testing of products in development. However, the lengthy turnaround time for detection of microbial contaminants when using a conventional sterility test is a bottleneck in this fast-paced environment. This study investigated an adenosine triphosphate-based bioluminescence rapid microbial method (RMM) for biologics samples, including monoclonal antibodies and hybridoma cell cultures. The results showed that the RMM allowed detection of antibody sample contaminants after only three days of incubation. In addition to being faster than the standard method, the RMM proved more reliable in detecting contaminants in cell culture samples with antibiotics. Since its initial evaluation, this RMM has accelerated biologics sterility testing across multiple projects at our site.
本研究将基于三磷酸腺苷（ATP）的生物发光快速微生物检测方法（RMM）与用于生物制品样品检测的传统无菌检测方法进行了比较。RMM基于在生长富集培养后对接种和未接种的微生物生长培养基样品中的ATP水平进行比较。本研究中合格的生物制品样品为重组单克隆抗体和杂交瘤细胞培养上清液。最初，这些样品背景ATP的批次间差异带来了重大挑战。评估了两种提高信噪比（阳性结果/背景ATP）的策略：基于酶的信号放大和通过培养基选择降低基于肉汤的噪声。在对RMM进行抗体和细胞培养样品的验证后，RMM还用于对多种纯化水源进行快速筛选。这种基于ATP的RMM在我们的发现研究站点对各种生物制品样品的常规检测中已证明具有极高价值，并在受污染样品的调查中发挥关键作用。生物制品研究实验室通常对处于研发阶段的产品进行无菌检测。然而，在这种快节奏的环境中，使用传统无菌检测方法检测微生物污染物的周转时间过长是一个瓶颈。本研究调查了一种用于生物制品样品（包括单克隆抗体和杂交瘤细胞培养物）的基于三磷酸腺苷的生物发光快速微生物检测方法（RMM）。结果表明，RMM仅需培养三天就能检测出抗体样品中的污染物。除了比标准方法更快之外，RMM在检测含抗生素的细胞培养样品中的污染物时也被证明更可靠。自最初评估以来，这种RMM已加速了我们站点多个项目的生物制品无菌检测。
