Department of Applied Chemistry, National Chiao Tung University, Hsinchu 300, Taiwan.
Department of Applied Chemistry, National Chiao Tung University, Hsinchu 300, Taiwan.
Biosens Bioelectron. 2017 Jun 15;92:410-416. doi: 10.1016/j.bios.2016.10.086. Epub 2016 Nov 1.
Ricin produced from the castor oil plant, Ricinus communis, is a well-known toxin. The toxin comprises A and B chains. Ricin A chain can cause toxicity by inhibiting protein synthesis, and ricin B can bind to the galactose ligand on the cell membrane of host cells. Inhalation or ingestion of ricin may even lead to death. Therefore, rapid and convenient sensing methods for detecting ricin in suspicious samples must be developed. In this study, we generated protein encapsulated gold nanoclusters (AuNCs@ew) with bright photoluminescence by using chicken egg white proteins as starting materials to react with aqueous tetrachloroaurate. The generated nanoclusters, which were mainly composed of chicken ovalbumin-encapsulated AuNCs, can recognize ricin B because of the presence of Galβ(1→4)GlcNAc ligands on chicken ovalbumin. The generated conjugates of AuNCs@ew and ricin B were heavy and readily settled down under centrifugation (13,000rpm, 60min). Thus, bright spots resulting from the conjugates at the bottom of the sample vials were easily visualized by the naked eye under ultraviolet light illumination. The limit of detection (LOD) was ~4.6µM. The LOD was reduced to ~400nM when fluorescence spectroscopy was used as the detection tool, while the LOD can be further improved to ~7.8nM when using matrix-assisted laser desorption/ionization mass spectrometry as the detection method. We also demonstrated the feasibility of using the proposed approach to selectively detect ricin B chain in complex samples.
蓖麻毒素是由大戟科植物蓖麻(Ricinus communis)产生的一种知名毒素。该毒素由 A 链和 B 链组成。蓖麻 A 链可通过抑制蛋白质合成而产生毒性,而蓖麻 B 链可与宿主细胞细胞膜上的半乳糖配体结合。吸入或摄入蓖麻毒素甚至可能导致死亡。因此,必须开发快速、方便的感测方法来检测可疑样本中的蓖麻毒素。在本研究中,我们使用鸡蛋白作为起始原料与水合四氯金酸反应,生成具有明亮光致发光的蛋白质包封金纳米团簇(AuNCs@ew)。所生成的纳米团簇主要由鸡卵清蛋白包封的 AuNCs 组成,由于鸡卵清蛋白上存在 Galβ(1→4)GlcNAc 配体,因此可以识别蓖麻 B 链。AuNCs@ew 和蓖麻 B 链生成的缀合物很重,在离心(13,000rpm,60min)下很容易沉降。因此,在紫外光照射下,很容易通过肉眼观察到位于样品小瓶底部的缀合物产生的亮斑。检测限(LOD)约为 4.6µM。当使用荧光光谱作为检测工具时,LOD 降低至约 400nM,而当使用基质辅助激光解吸/电离质谱作为检测方法时,LOD 可进一步提高至约 7.8nM。我们还证明了使用所提出的方法选择性地检测复杂样品中的蓖麻 B 链的可行性。