Nedelkov Dobrin, Nelson Randall W
Intrinsic Bioprobes Inc., Tempe, Arizona 85281, USA.
Appl Environ Microbiol. 2003 Sep;69(9):5212-5. doi: 10.1128/AEM.69.9.5212-5215.2003.
Detection of Staphylococcus enterotoxin B (SEB) by biomolecular interaction analysis mass spectrometry (BIA/MS) is presented in this work. The BIA/MS experiments were based on a surface plasmon resonance (SPR) MS immunoassay that detects affinity-captured SEB both via SPR and by means of exact and direct mass measurement by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Experiments were performed with standard samples and food samples to assess the BIA/MS limit of detection for SEB and to set the experimental parameters for proper quantitation. Single and double SPR referencing was performed to accurately estimate the amount of the bound toxin. Reproducible detection of 1 ng of SEB per ml, corresponding to affinity capture and MS analysis of approximately 500 amol of SEB, was readily achieved from both the standard and mushroom samples. A certain amount of SEB degradation was indicated by the signals in the mass spectra. The combination of MS with SPR-based methods of detection creates a unique approach capable of quantifying and qualitatively analyzing protein toxins from pathogenic organisms.
本文介绍了利用生物分子相互作用分析质谱法（BIA/MS）检测葡萄球菌肠毒素B（SEB）的方法。BIA/MS实验基于表面等离子体共振（SPR）质谱免疫分析法，该方法通过SPR以及基质辅助激光解吸/电离飞行时间（MALDI-TOF）质谱法进行精确直接的质量测量，来检测亲和捕获的SEB。使用标准样品和食品样品进行实验，以评估BIA/MS对SEB的检测限，并设置适当定量的实验参数。进行了单重和双重SPR参考，以准确估计结合毒素的量。从标准样品和蘑菇样品中都很容易实现每毫升1 ng SEB的可重复检测，这相当于对约500 amol的SEB进行亲和捕获和质谱分析。质谱图中的信号表明存在一定量的SEB降解。质谱与基于SPR的检测方法相结合，创造了一种能够对致病生物体中的蛋白质毒素进行定量和定性分析的独特方法。
