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<p>Here we report a novel platform for the detection of nucleocapsid (N) and receptor-binding domain (RBD) of spike (S) proteins of SARS-CoV-2 viruses using the surface plasmon resonance (SPR) technique. We demonstrate that the functionalization of SPR sensors with molecular 2D materials − 1&#xa0;nm thick carbon nanomembranes (CNMs) significantly enhances sensitivity. CNMs terminated with azide linker (N<sub>3</sub>-CNM) enable covalent bonding of SARS-CoV-2 antibodies for specific immobilization of the N- and S-proteins to the sensor surface. The successful and stable hierarchical functionalization is confirmed by multiparametric SPR measurements complemented with X-ray photoelectron spectroscopy and polarization modulation infrared reflection absorption spectroscopy. The obtained equilibrium dissociation constants (<i>K</i><sub><i>D</i></sub>) for the N-protein and the S-protein in the physiological buffer are 570 ± 50 pM and 22 ± 2 pM and the low detection limits (<i>LOD</i>s) are ~ 190 pM and ~ 10 pM, respectively. The high specificity of the developed sensors is shown via their negligible cross-reactivity with SARS-CoV-1 and MERS-CoV proteins. Finally, detection of SARS-CoV-2 proteins in nasopharyngeal swab samples with the <i>LOD</i> of ~ 40 pM is demonstrated. The proposed methodology enables the development of biosensors that cover clinically relevant range for the direct and immediate detection of SARS-CoV-2 without any amplification or labeling.</p>

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Highly sensitive and label-free detection of SARS-CoV-2 proteins via surface plasmon resonance using biofunctionalization with 1 nm thick carbon nanomembranes

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年8月25日

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  • Ghazaleh Eshaghi,
  • David Kaiser,
  • Hamid Reza Rasouli,
  • Rania Ennaciri,
  • Martha Frey,
  • Christof Neumann,
  • Dominik Gary,
  • Tobias Fischer,
  • Katrin Frankenfeld,
  • Andrey Turchanin

摘要

Here we report a novel platform for the detection of nucleocapsid (N) and receptor-binding domain (RBD) of spike (S) proteins of SARS-CoV-2 viruses using the surface plasmon resonance (SPR) technique. We demonstrate that the functionalization of SPR sensors with molecular 2D materials − 1 nm thick carbon nanomembranes (CNMs) significantly enhances sensitivity. CNMs terminated with azide linker (N3-CNM) enable covalent bonding of SARS-CoV-2 antibodies for specific immobilization of the N- and S-proteins to the sensor surface. The successful and stable hierarchical functionalization is confirmed by multiparametric SPR measurements complemented with X-ray photoelectron spectroscopy and polarization modulation infrared reflection absorption spectroscopy. The obtained equilibrium dissociation constants (KD) for the N-protein and the S-protein in the physiological buffer are 570 ± 50 pM and 22 ± 2 pM and the low detection limits (LODs) are ~ 190 pM and ~ 10 pM, respectively. The high specificity of the developed sensors is shown via their negligible cross-reactivity with SARS-CoV-1 and MERS-CoV proteins. Finally, detection of SARS-CoV-2 proteins in nasopharyngeal swab samples with the LOD of ~ 40 pM is demonstrated. The proposed methodology enables the development of biosensors that cover clinically relevant range for the direct and immediate detection of SARS-CoV-2 without any amplification or labeling.