tetano
Editor, Senior Moderator
Biosens Bioelectron
. 2022 Apr 8;209:114222.
doi: 10.1016/j.bios.2022.114222. Online ahead of print.
Performance of electrochemical immunoassays for clinical diagnostics of SARS-CoV-2 based on selective nucleocapsid N protein detection: Boron-doped diamond, gold and glassy carbon evaluation
Wioleta Białobrzeska[SUP] 1 [/SUP], Mateusz Ficek[SUP] 2 [/SUP], Bartłomiej Dec[SUP] 2 [/SUP], Silvio Osella[SUP] 3 [/SUP], Bartosz Trzaskowski[SUP] 3 [/SUP], Andres Jaramillo-Botero[SUP] 4 [/SUP], Mattia Pierpaoli[SUP] 2 [/SUP], Michał Rycewicz[SUP] 2 [/SUP], Yanina Dashkevich[SUP] 1 [/SUP], Tomasz Łęga[SUP] 1 [/SUP], Natalia Malinowska[SUP] 1 [/SUP], Zofia Cebula[SUP] 1 [/SUP], Daniel Bigus[SUP] 1 [/SUP], Daniel Firganek[SUP] 1 [/SUP], Ewelina Bięga[SUP] 1 [/SUP], Karolina Dziąbowska[SUP] 5 [/SUP], Mateusz Brodowski[SUP] 6 [/SUP], Marcin Kowalski[SUP] 6 [/SUP], Mirosława Panasiuk[SUP] 7 [/SUP], Beata Gromadzka[SUP] 7 [/SUP], Sabina Żołędowska[SUP] 8 [/SUP], Dawid Nidzworski[SUP] 5 [/SUP], Krzysztof Pyrć[SUP] 9 [/SUP], William A Goddard 3rd[SUP] 4 [/SUP], Robert Bogdanowicz[SUP] 10 [/SUP]
Affiliations
Abstract
The 21st century has already brought us a plethora of new threats related to viruses that emerge in humans after zoonotic transmission or drastically change their geographic distribution or prevalence. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first spotted at the end of 2019 to rapidly spread in southwest Asia and later cause a global pandemic, which paralyzes the world since then. We have designed novel immunosensors targeting conserved protein sequences of the N protein of SARS-CoV-2 based on lab-produced and purified anti-SARS-CoV-2 nucleocapsid antibodies that are densely grafted onto various surfaces (diamond/gold/glassy carbon). Titration of antibodies shows very strong reactions up to 1:72 900 dilution. Next, we showed the mechanism of interactions of our immunoassay with nucleocapsid N protein revealing molecular recognition by impedimetric measurements supported by hybrid modeling results with both density functional theory and molecular dynamics methods. Biosensors allowed for a fast (in less than 10 min) detection of SARS-CoV-2 virus with a limit of detection from 0.227 ng/ml through 0.334 ng/ml to 0.362 ng/ml for glassy carbon, boron-doped diamond, and gold surfaces, respectively. For all tested surfaces, we obtained a wide linear range of concentrations from 4.4 ng/ml to 4.4 pg/ml. Furthermore, our sensor leads to a highly specific response to SARS-CoV-2 clinical samples versus other upper respiratory tract viruses such as influenza, respiratory syncytial virus, or Epstein-Barr virus. All clinical samples were tested simultaneously on biosensors and real-time polymerase chain reactions.
. 2022 Apr 8;209:114222.
doi: 10.1016/j.bios.2022.114222. Online ahead of print.
Performance of electrochemical immunoassays for clinical diagnostics of SARS-CoV-2 based on selective nucleocapsid N protein detection: Boron-doped diamond, gold and glassy carbon evaluation
Wioleta Białobrzeska[SUP] 1 [/SUP], Mateusz Ficek[SUP] 2 [/SUP], Bartłomiej Dec[SUP] 2 [/SUP], Silvio Osella[SUP] 3 [/SUP], Bartosz Trzaskowski[SUP] 3 [/SUP], Andres Jaramillo-Botero[SUP] 4 [/SUP], Mattia Pierpaoli[SUP] 2 [/SUP], Michał Rycewicz[SUP] 2 [/SUP], Yanina Dashkevich[SUP] 1 [/SUP], Tomasz Łęga[SUP] 1 [/SUP], Natalia Malinowska[SUP] 1 [/SUP], Zofia Cebula[SUP] 1 [/SUP], Daniel Bigus[SUP] 1 [/SUP], Daniel Firganek[SUP] 1 [/SUP], Ewelina Bięga[SUP] 1 [/SUP], Karolina Dziąbowska[SUP] 5 [/SUP], Mateusz Brodowski[SUP] 6 [/SUP], Marcin Kowalski[SUP] 6 [/SUP], Mirosława Panasiuk[SUP] 7 [/SUP], Beata Gromadzka[SUP] 7 [/SUP], Sabina Żołędowska[SUP] 8 [/SUP], Dawid Nidzworski[SUP] 5 [/SUP], Krzysztof Pyrć[SUP] 9 [/SUP], William A Goddard 3rd[SUP] 4 [/SUP], Robert Bogdanowicz[SUP] 10 [/SUP]
Affiliations
- PMID: 35430407
- DOI: 10.1016/j.bios.2022.114222
Abstract
The 21st century has already brought us a plethora of new threats related to viruses that emerge in humans after zoonotic transmission or drastically change their geographic distribution or prevalence. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was first spotted at the end of 2019 to rapidly spread in southwest Asia and later cause a global pandemic, which paralyzes the world since then. We have designed novel immunosensors targeting conserved protein sequences of the N protein of SARS-CoV-2 based on lab-produced and purified anti-SARS-CoV-2 nucleocapsid antibodies that are densely grafted onto various surfaces (diamond/gold/glassy carbon). Titration of antibodies shows very strong reactions up to 1:72 900 dilution. Next, we showed the mechanism of interactions of our immunoassay with nucleocapsid N protein revealing molecular recognition by impedimetric measurements supported by hybrid modeling results with both density functional theory and molecular dynamics methods. Biosensors allowed for a fast (in less than 10 min) detection of SARS-CoV-2 virus with a limit of detection from 0.227 ng/ml through 0.334 ng/ml to 0.362 ng/ml for glassy carbon, boron-doped diamond, and gold surfaces, respectively. For all tested surfaces, we obtained a wide linear range of concentrations from 4.4 ng/ml to 4.4 pg/ml. Furthermore, our sensor leads to a highly specific response to SARS-CoV-2 clinical samples versus other upper respiratory tract viruses such as influenza, respiratory syncytial virus, or Epstein-Barr virus. All clinical samples were tested simultaneously on biosensors and real-time polymerase chain reactions.