Department of Otorhinolaryngology, Head and Neck Surgery, Section of Experimental Oncology and Nanomedicine (SEON), Else Kröner-Fresenius-Stiftung Professorship, Universitätsklinikum Erlangen, Erlangen, Germany.
Department of Experimental Physics 5 (Biophysics), Julius-Maximilians-University Würzburg, Würzburg, Germany.
Appl Microbiol Biotechnol. 2023 May;107(10):3329-3339. doi: 10.1007/s00253-023-12513-4. Epub 2023 Apr 15.
Pandemics like SARS-Cov-2 very frequently have their origin in different animals and in particular herds of camels could be a source of zoonotic diseases. This study took advantage on a highly sensitive and adaptable method for the fast and reliable detection of viral antibodies in camels using low-cost equipment. Magnetic nanoparticles (MNP) have high variability in their functionalization with different peptides and proteins. We confirm that 3-aminopropyl triethoxysilane (APTES)-coated MNP could be functionalized with viral proteins. The protein loading could be confirmed by simple loading controls using FACS-analysis (p < 0.05). Complementary combination of antigen and antibody yields in a significant signal increase could be proven by both FACS and COMPASS. However, COMPASS needs only a few seconds for the measurement. In COMPASS, the phase φ on selected critical point of the fifth higher harmonic (n = 5th). Here, positive sera display highly significant signal increase over the control or negative sera. Furthermore, a clear distinction could be made in antibody detection as an immune response to closely related viruses (SARS-CoV2 and MERS). Using modified MNPs along with COMPASS offers a fast and reliable method that is less cost intensive than current technologies and offers the possibility to be quickly adapted in case of new occurring viral infections. KEY POINTS: • COMPASS (critical offset magnetic particle spectroscopy) allows the fast detection of antibodies. • Magnetic nanoparticles can be adapted by exchange of the linked bait molecule. • Antibodies could be detected in camel sera without washing steps within seconds.
像 SARS-CoV-2 这样的大流行病毒经常起源于不同的动物,特别是骆驼群可能是人畜共患病的源头。本研究利用一种高度敏感和适应性强的方法,利用低成本设备快速可靠地检测骆驼中的病毒抗体。磁性纳米颗粒(MNP)在其与不同肽和蛋白质的功能化方面具有高度的可变性。我们证实,3-氨丙基三乙氧基硅烷(APTES)涂覆的 MNP 可以用病毒蛋白进行功能化。通过使用 FAC 分析的简单加载对照(p < 0.05)可以确认蛋白质加载。通过 FACS 和 COMPASS 都可以证明抗原和抗体的互补组合可以产生显著的信号增加。然而,COMPASS 仅需几秒钟即可进行测量。在 COMPASS 中,相位 φ 位于第五个更高谐波的选定临界点(n = 5th)上。在这里,阳性血清显示出比对照或阴性血清显著增加的信号。此外,还可以在抗体检测中做出明确区分,因为这是对密切相关的病毒(SARS-CoV2 和 MERS)的免疫反应。使用经过修饰的 MNPs 结合 COMPASS 提供了一种快速可靠的方法,比当前技术成本更低,并为新出现的病毒感染提供了快速适应的可能性。要点:• COMPASS(关键偏移磁性粒子光谱法)允许快速检测抗体。• 可以通过交换连接的诱饵分子来适应磁性纳米颗粒。• 无需洗涤步骤,可在几秒钟内在骆驼血清中检测到抗体。