Digital and Analog Detection of SARS-CoV-2 Nucleocapsid Protein
via an Upconversion-Linked Immunosorbent Assay

J 2023

Digital and Analog Detection of SARS-CoV-2 Nucleocapsid Protein via an Upconversion-Linked Immunosorbent Assay

BRANDMEIER, Julian, Natalia JURGA, Tomasz GRZYB, Antonín HLAVÁČEK, Radka OBOŘILOVÁ et. al.

Basic information

Original name

Digital and Analog Detection of SARS-CoV-2 Nucleocapsid Protein via an Upconversion-Linked Immunosorbent Assay

Authors

BRANDMEIER, Julian (276 Germany, belonging to the institution), Natalia JURGA, Tomasz GRZYB (616 Poland), Antonín HLAVÁČEK (203 Czech Republic), Radka OBOŘILOVÁ (203 Czech Republic, belonging to the institution), Petr SKLÁDAL (203 Czech Republic, belonging to the institution), Zdeněk FARKA (203 Czech Republic, belonging to the institution) and Hans-Heiner GORRIS (276 Germany, guarantor, belonging to the institution)

Edition

Analytical Chemistry, Washington, DC, American Chemical Society, 2023, 0003-2700

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10406 Analytical chemistry

Country of publisher

United States of America

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 7.400 in 2022

RIV identification code

RIV/00216224:14310/23:00130445

Organization unit

Faculty of Science

DOI

http://dx.doi.org/10.1021/acs.analchem.2c05670

UT WoS

000939526700001

Keywords in English

Covid-19; SARS-CoV-2; immunoassay; photon-upconversion nanoparticle; ULISA; digital detection

Abstract

V originále

The COVID-19 crisis requires fast and highly sensitive tests for the early stage detection of the SARS-CoV-2 virus. For detecting the nucleocapsid protein (N protein), the most abundant viral antigen, we have employed upconversion nanoparticles that emit short-wavelength light under near-infrared excitation (976 nm). The anti-Stokes emission avoids autofluorescence and light scattering and thus enables measurements without optical background interference. The sandwich upconversion-linked immunosorbent assay (ULISA) can be operated both in a conventional analog mode and in a digital mode based on counting individual immune complexes. We have investigated how different antibody combinations affect the detection of the wildtype N protein and the detection of SARS-CoV-2 (alpha variant) in lysed culture fluid via the N protein. The ULISA yielded a limit of detection (LOD) of 1.3 pg/mL (27 fM) for N protein detection independent of the analog or digital readout, which is approximately 3 orders of magnitude more sensitive than conventional enzyme-linked immunosorbent assays or commercial lateral flow assays for home testing. In the case of SARS-CoV-2, the digital ULISA additionally improved the LOD by a factor of 10 compared to the analog readout.

Links

GA21-03156S, research and development project

Name: Foton-upkonverzní značky pro mikrofluidní jednomolekulové imunostanovení proteinových biomarkerů

Investor: Czech Science Foundation

LM2018127, research and development project

Name: Česká infrastruktura pro integrativní strukturní biologii (Acronym: CIISB)

Investor: Ministry of Education, Youth and Sports of the CR

90127, large research infrastructures

Name: CIISB II

Citovat

BRANDMEIER, Julian, Natalia JURGA, Tomasz GRZYB, Antonín HLAVÁČEK, Radka OBOŘILOVÁ, Petr SKLÁDAL, Zdeněk FARKA and Hans-Heiner GORRIS. Digital and Analog Detection of SARS-CoV-2 Nucleocapsid Protein via an Upconversion-Linked Immunosorbent Assay. Analytical Chemistry. Washington, DC: American Chemical Society, 2023, vol. 95, No 10, p. 4753-4759. ISSN 0003-2700. Available from: https://dx.doi.org/10.1021/acs.analchem.2c05670.

@article{2267281,
   author = {Brandmeier, Julian and Jurga, Natalia and Grzyb, Tomasz and Hlaváček, Antonín and Obořilová, Radka and Skládal, Petr and Farka, Zdeněk and Gorris, HansandHeiner},
   article_location = {Washington, DC},
   article_number = {10},
   doi = {http://dx.doi.org/10.1021/acs.analchem.2c05670},
   keywords = {Covid-19; SARS-CoV-2; immunoassay; photon-upconversion nanoparticle; ULISA; digital detection},
   language = {eng},
   issn = {0003-2700},
   journal = {Analytical Chemistry},
   title = {Digital and Analog Detection of SARS-CoV-2 Nucleocapsid Protein via an Upconversion-Linked Immunosorbent Assay},
   url = {https://pubs.acs.org/doi/10.1021/acs.analchem.2c05670},
   volume = {95},
   year = {2023}
}

TY  - JOUR
ID  - 2267281
AU  - Brandmeier, Julian - Jurga, Natalia - Grzyb, Tomasz - Hlaváček, Antonín - Obořilová, Radka - Skládal, Petr - Farka, Zdeněk - Gorris, Hans-Heiner
PY  - 2023
TI  - Digital and Analog Detection of SARS-CoV-2 Nucleocapsid Protein via an Upconversion-Linked Immunosorbent Assay
JF  - Analytical Chemistry
VL  - 95
IS  - 10
SP  - 4753-4759
EP  - 4753-4759
PB  - American Chemical Society
SN  - 00032700
KW  - Covid-19
KW  - SARS-CoV-2
KW  - immunoassay
KW  - photon-upconversion nanoparticle
KW  - ULISA
KW  - digital detection
UR  - https://pubs.acs.org/doi/10.1021/acs.analchem.2c05670
N2  - The COVID-19 crisis requires fast and highly sensitive tests for the early stage detection of the SARS-CoV-2 virus. For detecting the nucleocapsid protein (N protein), the most abundant viral antigen, we have employed upconversion nanoparticles that emit short-wavelength light under near-infrared excitation (976 nm). The anti-Stokes emission avoids autofluorescence and light scattering and thus enables measurements without optical background interference. The sandwich upconversion-linked immunosorbent assay (ULISA) can be operated both in a conventional analog mode and in a digital mode based on counting individual immune complexes. We have investigated how different antibody combinations affect the detection of the wildtype N protein and the detection of SARS-CoV-2 (alpha variant) in lysed culture fluid via the N protein. The ULISA yielded a limit of detection (LOD) of 1.3 pg/mL (27 fM) for N protein detection independent of the analog or digital readout, which is approximately 3 orders of magnitude more sensitive than conventional enzyme-linked immunosorbent assays or commercial lateral flow assays for home testing. In the case of SARS-CoV-2, the digital ULISA additionally improved the LOD by a factor of 10 compared to the analog readout.
ER  -

BRANDMEIER, Julian, Natalia JURGA, Tomasz GRZYB, Antonín HLAVÁČEK, Radka OBOŘILOVÁ, Petr SKLÁDAL, Zdeněk FARKA and Hans-Heiner GORRIS. Digital and Analog Detection of SARS-CoV-2 Nucleocapsid Protein via an Upconversion-Linked Immunosorbent Assay. \textit{Analytical Chemistry}. Washington, DC: American Chemical Society, 2023, vol.~95, No~10, p.~4753-4759. ISSN~0003-2700. Available from: https://dx.doi.org/10.1021/acs.analchem.2c05670.

Detailed Information on Publication Record