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Single Domain Antibodies for the Detection of Ricin Using Silicon Photonic Microring Resonator Arrays

机译:使用硅光子微环谐振器阵列检测蓖麻毒素的单域抗体

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Ricin is a lethal protein toxin derived from the castor bean plant. Given its notorious history as a biowarfare agent and homicidal weapon, ricin has been classified as a category B bioterrorism agent. Current ricin detection methods based on immunoassays lack the required sensitivity and specificity for many homeland security surveillance applications. Importantly, many conventional antibody-based methodologies are unable to distinguish ricin from RCA 120, a nontoxic protein also found in the castor bean plant. Single domain antibodies (sdAbs), which are recombinantly derived from immunized llamas, are known to have high affinities for ricin A or B chains and low cross-reactivity with RCA 120. Herein, we demonstrate the use of silicon photonic microring resonators for antibody affinity profiling and one-step ricin detection at concentrations down to 300 pM using a 15 min, label-free assay format. These sdAbs were also simultaneously compared with a commercial anti-RCA IgG antibody in a multicapture agent, single target immunoassay using arrays of microrings, which allowed direct comparison of sensitivity and specificity. A selected sdAb was also found to exhibit outstanding specificity against another biotoxin, saporin, which has mechanism of action similar to ricin. Given the rapidity, scalability, and multiplexing capability of this silicon-based technology, this work represents a step toward using microring resonator arrays for the sensitive and specific detection of biowarfare agents.
机译:蓖麻毒素是一种源自蓖麻子植物的致命蛋白质毒素。蓖麻毒素以其作为生物战剂和杀人武器的臭名昭著的历史而被列为B类生物恐怖主义剂。当前基于免疫测定的蓖麻毒蛋白检测方法缺乏许多国土安全监视应用所需的敏感性和特异性。重要的是,许多传统的基于抗体的方法无法将蓖麻毒素与RCA 120区别开,RCA 120是在蓖麻子植物中也发现的一种无毒蛋白质。单域抗体(sdAbs)重组衍生自免疫的美洲驼,已知对蓖麻毒蛋白A或B链具有高亲和力,并且与RCA 120的交叉反应性较低。在此,我们证明了利用硅光子微环共振器实现抗体亲和力使用15分钟,无标记的测定形式进行低至300 pM浓度的分析和一步一步蓖麻毒素检测。这些sdAb还与多捕获剂中的市售抗RCA IgG抗体同时进行了比较,使用微环阵列进行了单靶免疫测定,从而可以直接比较灵敏度和特异性。还发现一种选定的sdAb对另一种生物毒素-沙泊菌素(saporin)表现出杰出的特异性,其作用机理与蓖麻毒蛋白相似。考虑到这种基于硅的技术的快速性,可扩展性和多路复用能力,这项工作代表了朝着使用微环谐振器阵列进行生物战剂的灵敏和特异检测的一步。

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