Kalb Suzanne R, Schieltz David M, Becher François, Astot Crister, Fredriksson Sten-Åke, Barr John R
Centers for Disease Control and Prevention, 4770 Buford Hwy NE, Atlanta, GA 30341, USA.
Service de Pharmacologie et d'Immunoanalyse, Institut de Biologie et de Technologies de Saclay (iBiTec-S), Commissariat à l'Énergie Atomique et aux Énergies Alternatives (CEA), 91191 Gif-sur-Yvette, France.
Toxins (Basel). 2015 Nov 25;7(12):4881-94. doi: 10.3390/toxins7124854.
Ricin is a protein toxin produced by the castor bean plant (Ricinus communis) together with a related protein known as R. communis agglutinin (RCA120). Mass spectrometric (MS) assays have the capacity to unambiguously identify ricin and to detect ricin's activity in samples with complex matrices. These qualitative and quantitative assays enable detection and differentiation of ricin from the less toxic RCA120 through determination of the amino acid sequence of the protein in question, and active ricin can be monitored by MS as the release of adenine from the depurination of a nucleic acid substrate. In this work, we describe the application of MS-based methods to detect, differentiate and quantify ricin and RCA120 in nine blinded samples supplied as part of the EQuATox proficiency test. Overall, MS-based assays successfully identified all samples containing ricin or RCA120 with the exception of the sample spiked with the lowest concentration (0.414 ng/mL). In fact, mass spectrometry was the most successful method for differentiation of ricin and RCA120 based on amino acid determination. Mass spectrometric methods were also successful at ranking the functional activities of the samples, successfully yielding semi-quantitative results. These results indicate that MS-based assays are excellent techniques to detect, differentiate, and quantify ricin and RCA120 in complex matrices.
蓖麻毒素是一种由蓖麻植株(蓖麻)产生的蛋白质毒素,同时还产生一种相关蛋白,即蓖麻凝集素(RCA120)。质谱(MS)分析能够明确鉴定蓖麻毒素,并能在含有复杂基质的样品中检测其活性。这些定性和定量分析通过确定相关蛋白质的氨基酸序列,实现了从毒性较低的RCA120中检测和区分蓖麻毒素,并且作为核酸底物脱嘌呤过程中腺嘌呤的释放,活性蓖麻毒素可以通过质谱进行监测。在这项工作中,我们描述了基于质谱的方法在检测、区分和定量九份作为EQuATox能力验证测试一部分提供的盲样中的蓖麻毒素和RCA120方面的应用。总体而言,基于质谱的分析成功鉴定了所有含有蓖麻毒素或RCA120的样品,但加样浓度最低(0.414 ng/mL)的样品除外。事实上,基于氨基酸测定,质谱是区分蓖麻毒素和RCA120最成功的方法。质谱方法在对样品的功能活性进行排序方面也很成功,成功得出了半定量结果。这些结果表明,基于质谱的分析是在复杂基质中检测、区分和定量蓖麻毒素和RCA120的优秀技术。