DEVELOPMENT AND VALIDATION OF A BIOASSAY FOR INTERLEUKIN-2
DEVELOPMENT AND VALIDATION OF A BIOASSAY FOR INTERLEUKIN-2
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DOI：
10.1016/0731-7085(92)80079-3
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发表时间：
1992-08-01
影响因子：
3.4
通讯作者：
SJODIN, L
中科院分区：
文献类型：
--
作者：
HAMMERLING, U;HENNINGSSON, AC;SJODIN, L
建立了一种基于刺激CTLL细胞增殖的IL-2活性测定的可靠而精确的方法。将细胞与不同浓度的IL-2在微量滴定板中孵育24小时。通过阅读甲瓒的光密度，在平板-阅读分光光度计上测量刺激作用，甲瓒由来自3-（4，5-二甲基噻唑-2-基）-2，5-二苯基四唑溴化盐（MTT）的活细胞产生。生物测定设计为四剂量平行线试验，符合药典对测定有效性的要求，一组四个实验的批间相对标准偏差（RSD）为2.6%。采用人IL-2的国际标准品和重组DNA衍生IL-2的参比试剂进行效价测定。发现该方法适用于IL-2的药物制剂的效力评估。
A reliable and precise method for the determination of IL-2 activity, based on stimulation of CTLL cell proliferation, was developed. Cells were incubated with different concentrations of IL-2 for 24 h in microtiter plates. The stimulatory effect was measured on a plate-reading spectrophotometer by reading the optical density of formazan, which is produced by viable cells from 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The bioassay was designed as a four-dose parallel line test, fulfilling pharmacopoeial requirements for assay validity, and the inter-assay relative standard deviation (RSD) for a group of four experiments was 2.6%. The International Standard for human IL-2 and the Reference Reagent for Recombinant DNA-derived IL-2 were employed for potency determinations. The method was found suitable for potency assessments of pharmaceutical formulations of IL-2.