Hale M L
Toxinology Division, United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, MD 21702 5011, USA.
Pharmacol Toxicol. 2001 May;88(5):255-60.
A microtiter assay was developed to quickly measure the biological activity of ricin or other ribosome-inactivating proteins. Nuclease-treated rabbit reticulocyte lysate containing luciferase mRNA was used to measure toxin activity via inhibition of protein synthesis. The relative biological activity was determined by comparing luminescence levels in treated samples versus those of untreated controls. The amount of luciferase translated, as measured by luminescence, was inversely proportional to the toxin concentration. Linear dose response curves were generated for both class I and class II toxins. When compared to normal serum controls, specific antibody against ricin effectively inhibited ricin activity. The assay is suitable for efficient in vitro screening of therapeutics, as well as for identifying samples containing ribosome-inactivating proteins.
开发了一种微量滴定法,用于快速测量蓖麻毒素或其他核糖体失活蛋白的生物活性。使用经核酸酶处理的含有荧光素酶mRNA的兔网织红细胞裂解物,通过抑制蛋白质合成来测量毒素活性。通过比较处理样品与未处理对照的发光水平来确定相对生物活性。通过发光测量的荧光素酶翻译量与毒素浓度成反比。为I类和II类毒素均生成了线性剂量反应曲线。与正常血清对照相比,抗蓖麻毒素的特异性抗体有效抑制了蓖麻毒素活性。该测定法适用于治疗药物的高效体外筛选,以及鉴定含有核糖体失活蛋白的样品。