Pellett Sabine, Tepp William H, Toth Stephen I, Johnson Eric A
Department of Bacteriology, University of Wisconsin, Madison, Madison, WI 53706, USA.
J Pharmacol Toxicol Methods. 2010 May-Jun;61(3):304-10. doi: 10.1016/j.vascn.2010.01.003. Epub 2010 Jan 25.
Botulinum neurotoxin (BoNT) type A is increasingly used in humans for pharmaceutical and cosmetic purposes. Currently, the standard assay used to determine potency of clinical samples, and the only assay approved by the FDA, is the in vivo mouse bioassay (MBA). However, due to several drawbacks of this assay (relatively large error, high cost, no standardization, requirement of high technical expertise, and use of large numbers of mice), there is an increasing need to replace this assay. A cell-based assay using primary rat spinal cord cells (RSC assay) has been previously reported to sensitively detect purified botulinum neurotoxin type A, and requires all biological properties of the toxin for detection.
This study presents data on quantitative detection of potency of purified BoNT/A by a cell-based assay, using primary rat spinal cord cells (RSC assay). The sensitivity and error rate of the RSC assay was directly compared to the currently used mouse bioassay by repeated testing of the same purified BoNT/A sample by both assays. In addition, the potency of several samples of purified BoNT/A of unknown activity was determined in parallel by RSC assay and MBA.
The results indicate sensitivity of the RSC assay similar to the mouse bioassay, high reproducibility, and a lower error rate than the mouse bioassay. Direct comparison of potency determination of several purified BoNT/A samples by RSC assay and MBA resulted in very similar values, indicating very good correlation.
These data support the use of a cell-based assay for potency determination of purified BoNT/A as an alternative to the mouse bioassay.
A型肉毒杆菌神经毒素（BoNT）在人类医药和美容领域的应用日益广泛。目前，用于测定临床样本效力的标准检测方法，也是美国食品药品监督管理局（FDA）唯一批准的检测方法，是体内小鼠生物测定法（MBA）。然而，由于该检测方法存在诸多缺点（相对误差大、成本高、缺乏标准化、需要高技术专业知识以及使用大量小鼠），越来越需要替代该检测方法。此前有报道称，一种使用原代大鼠脊髓细胞的基于细胞的检测方法（RSC检测法）能够灵敏地检测纯化的A型肉毒杆菌神经毒素，且检测需要毒素的所有生物学特性。
本研究展示了使用原代大鼠脊髓细胞的基于细胞的检测方法（RSC检测法）对纯化的BoNT/A效力进行定量检测的数据。通过对同一纯化的BoNT/A样本进行两种检测方法的重复测试，将RSC检测法的灵敏度和误差率与目前使用的小鼠生物测定法直接进行比较。此外，通过RSC检测法和MBA平行测定了几个活性未知的纯化BoNT/A样本的效力。
结果表明，RSC检测法的灵敏度与小鼠生物测定法相似，具有高重现性，且误差率低于小鼠生物测定法。通过RSC检测法和MBA对几个纯化的BoNT/A样本的效力测定进行直接比较，得到了非常相似的值，表明相关性非常好。
这些数据支持使用基于细胞的检测方法来测定纯化的BoNT/A的效力，作为小鼠生物测定法的替代方法。
