{"title":"开发基于金纳米粒子的新型诊断原型,用于体内检测印度红蝎(Mesobuthus tamulus)毒液","authors":"","doi":"10.1016/j.toxcx.2024.100203","DOIUrl":null,"url":null,"abstract":"<div><p>Indian red scorpion <em>Mesobuthus tamulus</em> is responsible for substantial mortality in India and Sri Lanka; however, no specific diagnostic method is available to detect the venom of this scorpion in envenomed plasma or body fluid. Therefore, we have proposed a novel, simple, and rapid method for detecting <em>M. tamulus</em> venom (MTV) in the plasma of envenomed animals using polyclonal antibodies (PAb) raised against three modified custom peptides representing the antigenic epitopes of K<sup>+</sup> (Tamapin) and Na<sup>+</sup> (α-neurotoxin) channel toxins, the two major MTV toxins identified by proteomic analysis. The optimum PAb formulation containing PAb 1, 2, and 3 in proportion (1:1:1, w/w/w) acted synergistically, demonstrating significantly higher immunological recognition of MTV than anti-scorpion antivenom (developed against native toxins) and individual antibodies against peptide immunogens. The PAb formulation could detect MTV optimally in envenomed rat plasma (intravenous and subcutaneous routes) at 30–60 min post-injection. The acetonitrile precipitation method developed in this study to augment the MTV detection sensitivity enriched the low molecular mass peptide toxins in envenomed rat plasma, which was ascertained by mass spectrometry analysis. The gold nanoparticles conjugated PAb formulation, characterised by biophysical techniques such as Fourier transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM), demonstrated their interaction with low molecular mass MTV peptide toxins in envenomed rat plasma. This interaction results in the accumulation of the gold nanoparticles, thus leading to signal change in absorbance spectra that can be discerned within 10 min. From a standard curve of MTV spiked plasma, the quantity of MTV in envenomed rat plasma could be determined by gold nanoparticle-PAb formulation conjugate.</p></div>","PeriodicalId":37124,"journal":{"name":"Toxicon: X","volume":null,"pages":null},"PeriodicalIF":3.6000,"publicationDate":"2024-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2590171024000201/pdfft?md5=8238eec18bed45328ca818b6d09a6e67&pid=1-s2.0-S2590171024000201-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Development of a gold nanoparticle-based novel diagnostic prototype for in vivo detection of Indian red scorpion (Mesobuthus tamulus) venom\",\"authors\":\"\",\"doi\":\"10.1016/j.toxcx.2024.100203\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Indian red scorpion <em>Mesobuthus tamulus</em> is responsible for substantial mortality in India and Sri Lanka; however, no specific diagnostic method is available to detect the venom of this scorpion in envenomed plasma or body fluid. Therefore, we have proposed a novel, simple, and rapid method for detecting <em>M. tamulus</em> venom (MTV) in the plasma of envenomed animals using polyclonal antibodies (PAb) raised against three modified custom peptides representing the antigenic epitopes of K<sup>+</sup> (Tamapin) and Na<sup>+</sup> (α-neurotoxin) channel toxins, the two major MTV toxins identified by proteomic analysis. The optimum PAb formulation containing PAb 1, 2, and 3 in proportion (1:1:1, w/w/w) acted synergistically, demonstrating significantly higher immunological recognition of MTV than anti-scorpion antivenom (developed against native toxins) and individual antibodies against peptide immunogens. The PAb formulation could detect MTV optimally in envenomed rat plasma (intravenous and subcutaneous routes) at 30–60 min post-injection. The acetonitrile precipitation method developed in this study to augment the MTV detection sensitivity enriched the low molecular mass peptide toxins in envenomed rat plasma, which was ascertained by mass spectrometry analysis. The gold nanoparticles conjugated PAb formulation, characterised by biophysical techniques such as Fourier transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM), demonstrated their interaction with low molecular mass MTV peptide toxins in envenomed rat plasma. This interaction results in the accumulation of the gold nanoparticles, thus leading to signal change in absorbance spectra that can be discerned within 10 min. 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引用次数: 0
摘要
印度红蝎(Mesobuthus tamulus)是造成印度和斯里兰卡大量死亡的罪魁祸首;然而,目前还没有特定的诊断方法来检测被毒死动物血浆或体液中的这种蝎子的毒液。因此,我们提出了一种新颖、简单、快速的方法,利用针对三种经修饰的定制肽的多克隆抗体(PAb)来检测被毒杀动物血浆中的蝎毒(MTV),这三种肽分别代表了 K+(Tamapin)和 Na+(α-神经毒素)通道毒素的抗原表位,而这两种毒素是通过蛋白质组分析确定的两种主要的 MTV 毒素。含有 PAb 1、2 和 3 的最佳 PAb 配方按比例(1:1:1, w/w/w)协同作用,对 MTV 的免疫识别率明显高于抗蝎抗血清(针对原生毒素开发)和针对肽免疫原的单个抗体。PAb 制剂可在注射后 30-60 分钟内在被毒死的大鼠血浆(静脉注射和皮下注射)中检测出 MTV。本研究开发的乙腈沉淀法提高了 MTV 的检测灵敏度,富集了被毒杀大鼠血浆中的低分子质量多肽毒素,这一点已通过质谱分析得到证实。通过傅立叶变换红外光谱(FTIR)和透射电子显微镜(TEM)等生物物理技术对金纳米粒子共轭 PAb 制剂进行表征,结果表明它们与被毒杀大鼠血浆中的低分子质量 MTV 多肽毒素发生了相互作用。这种相互作用会导致金纳米粒子的积累,从而导致吸光度光谱的信号变化,这种变化可在 10 分钟内分辨出来。根据添加了 MTV 的血浆的标准曲线,可通过金纳米粒子-PAb 制剂共轭物确定被毒杀大鼠血浆中 MTV 的含量。
Development of a gold nanoparticle-based novel diagnostic prototype for in vivo detection of Indian red scorpion (Mesobuthus tamulus) venom
Indian red scorpion Mesobuthus tamulus is responsible for substantial mortality in India and Sri Lanka; however, no specific diagnostic method is available to detect the venom of this scorpion in envenomed plasma or body fluid. Therefore, we have proposed a novel, simple, and rapid method for detecting M. tamulus venom (MTV) in the plasma of envenomed animals using polyclonal antibodies (PAb) raised against three modified custom peptides representing the antigenic epitopes of K+ (Tamapin) and Na+ (α-neurotoxin) channel toxins, the two major MTV toxins identified by proteomic analysis. The optimum PAb formulation containing PAb 1, 2, and 3 in proportion (1:1:1, w/w/w) acted synergistically, demonstrating significantly higher immunological recognition of MTV than anti-scorpion antivenom (developed against native toxins) and individual antibodies against peptide immunogens. The PAb formulation could detect MTV optimally in envenomed rat plasma (intravenous and subcutaneous routes) at 30–60 min post-injection. The acetonitrile precipitation method developed in this study to augment the MTV detection sensitivity enriched the low molecular mass peptide toxins in envenomed rat plasma, which was ascertained by mass spectrometry analysis. The gold nanoparticles conjugated PAb formulation, characterised by biophysical techniques such as Fourier transform infrared spectroscopy (FTIR) and transmission electron microscopy (TEM), demonstrated their interaction with low molecular mass MTV peptide toxins in envenomed rat plasma. This interaction results in the accumulation of the gold nanoparticles, thus leading to signal change in absorbance spectra that can be discerned within 10 min. From a standard curve of MTV spiked plasma, the quantity of MTV in envenomed rat plasma could be determined by gold nanoparticle-PAb formulation conjugate.