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.

Základní údaje

Originální název

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

Autoři

BRANDMEIER, Julian (276 Německo, domácí), Natalia JURGA, Tomasz GRZYB (616 Polsko), Antonín HLAVÁČEK (203 Česká republika), Radka OBOŘILOVÁ (203 Česká republika, domácí), Petr SKLÁDAL (203 Česká republika, domácí), Zdeněk FARKA (203 Česká republika, domácí) a Hans-Heiner GORRIS (276 Německo, garant, domácí)

Vydání

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

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10406 Analytical chemistry

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 7.400 v roce 2022

Kód RIV

RIV/00216224:14310/23:00130445

Organizační jednotka

Přírodovědecká fakulta

DOI

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

UT WoS

000939526700001

Klíčová slova anglicky

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

Štítky

CF NANO, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 17. 10. 2024 17:49, Mgr. Adéla Pešková

Anotace

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.

Návaznosti

GA21-03156S, projekt VaV

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

Investor: Grantová agentura ČR, Foton-upkonverzní značky pro mikrofluidní jednomolekulové imunostanovení proteinových biomarkerů

LM2018127, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii (Akronym: CIISB)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Czech Infrastructure for Integrative Structural Biology

90127, velká výzkumná infrastruktura

Název: CIISB II

Citovat

BRANDMEIER, Julian, Natalia JURGA, Tomasz GRZYB, Antonín HLAVÁČEK, Radka OBOŘILOVÁ, Petr SKLÁDAL, Zdeněk FARKA a 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, roč. 95, č. 10, s. 4753-4759. ISSN 0003-2700. Dostupné z: 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 a 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, roč.~95, č.~10, s.~4753-4759. ISSN~0003-2700. Dostupné z: https://dx.doi.org/10.1021/acs.analchem.2c05670.

Podrobný výpis o publikaci