tetano
Editor, Senior Moderator
ACS Appl Mater Interfaces
. 2023 Jul 25.
doi: 10.1021/acsami.3c05900. Online ahead of print. Development of Nanobody-Displayed Whole-Cell Biosensors for the Colorimetric Detection of SARS-CoV-2
Yawen He[SUP] 1 [/SUP], Zhiyuan Xu[SUP] 1 [/SUP], Tom Kasputis[SUP] 1 [/SUP], Xue Zhao[SUP] 1 [/SUP], Itati Ibañez[SUP] 2 [/SUP], Florencia Pavan[SUP] 2 [/SUP], Marina Bok[SUP] 3 [/SUP], Juan Pablo Malito[SUP] 3 [/SUP], Viviana Parreno[SUP] 3 4 [/SUP], Lijuan Yuan[SUP] 4 [/SUP], R Clay Wright[SUP] 1 [/SUP], Juhong Chen[SUP] 1 [/SUP]
Affiliations
The accurate and effective detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is essential to preventing the spread of infectious diseases and ensuring human health. Herein, a nanobody-displayed whole-cell biosensor was developed for colorimetric detection of SARS-CoV-2 spike proteins. Serving as bioreceptors, yeast surfaces were genetically engineered to display SARS-CoV-2 binding of llama-derived single-domain antibodies (nanobodies) with high capture efficiency, facilitating the concentration and purification of SARS-CoV-2. Gold nanoparticles (AuNPs) employed as signal transductions were functionalized with horseradish peroxidase (HRP) and anti-SARS monoclonal antibodies to enhance the detection sensitivity. In the presence of SARS-CoV-2 spike proteins, the sandwiched binding will be formed by linking engineered yeast, SARS-CoV-2 spike proteins, and reporter AuNPs. The colorimetric signal was generated by the enzymatic reaction of HRP and its corresponding colorimetric substrate/chromogen system. At the optimal conditions, the developed whole-cell biosensor enables the sensitive detection of SARS-CoV-2 spike proteins in a linear range from 0.01 to 1 μg/mL with a limit of detection (LOD) of 0.037 μg/mL (about 4 × 10[SUP]8[/SUP] virion particles/mL). Furthermore, the whole-cell biosensor was demonstrated to detect the spike protein of different SARS-CoV-2 variants in human serum, providing new possibilities for the detection of future SARS-CoV-2 variants.
Keywords: SARS-CoV-2; colorimetric detection; engineered yeast; gold nanoparticle; nanobody; whole-cell biosensor.
. 2023 Jul 25.
doi: 10.1021/acsami.3c05900. Online ahead of print. Development of Nanobody-Displayed Whole-Cell Biosensors for the Colorimetric Detection of SARS-CoV-2
Yawen He[SUP] 1 [/SUP], Zhiyuan Xu[SUP] 1 [/SUP], Tom Kasputis[SUP] 1 [/SUP], Xue Zhao[SUP] 1 [/SUP], Itati Ibañez[SUP] 2 [/SUP], Florencia Pavan[SUP] 2 [/SUP], Marina Bok[SUP] 3 [/SUP], Juan Pablo Malito[SUP] 3 [/SUP], Viviana Parreno[SUP] 3 4 [/SUP], Lijuan Yuan[SUP] 4 [/SUP], R Clay Wright[SUP] 1 [/SUP], Juhong Chen[SUP] 1 [/SUP]
Affiliations
- PMID: 37489943
- DOI: 10.1021/acsami.3c05900
The accurate and effective detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is essential to preventing the spread of infectious diseases and ensuring human health. Herein, a nanobody-displayed whole-cell biosensor was developed for colorimetric detection of SARS-CoV-2 spike proteins. Serving as bioreceptors, yeast surfaces were genetically engineered to display SARS-CoV-2 binding of llama-derived single-domain antibodies (nanobodies) with high capture efficiency, facilitating the concentration and purification of SARS-CoV-2. Gold nanoparticles (AuNPs) employed as signal transductions were functionalized with horseradish peroxidase (HRP) and anti-SARS monoclonal antibodies to enhance the detection sensitivity. In the presence of SARS-CoV-2 spike proteins, the sandwiched binding will be formed by linking engineered yeast, SARS-CoV-2 spike proteins, and reporter AuNPs. The colorimetric signal was generated by the enzymatic reaction of HRP and its corresponding colorimetric substrate/chromogen system. At the optimal conditions, the developed whole-cell biosensor enables the sensitive detection of SARS-CoV-2 spike proteins in a linear range from 0.01 to 1 μg/mL with a limit of detection (LOD) of 0.037 μg/mL (about 4 × 10[SUP]8[/SUP] virion particles/mL). Furthermore, the whole-cell biosensor was demonstrated to detect the spike protein of different SARS-CoV-2 variants in human serum, providing new possibilities for the detection of future SARS-CoV-2 variants.
Keywords: SARS-CoV-2; colorimetric detection; engineered yeast; gold nanoparticle; nanobody; whole-cell biosensor.