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; Natalia JURGA; Tomasz GRZYB; Antonín HLAVÁČEK; Radka OBOŘILOVÁ; Petr SKLÁDAL; Zdeněk FARKA a Hans-Heiner GORRIS

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: 6.800

Kód RIV

RIV/00216224:14310/23:00130445

Organizační jednotka

Přírodovědecká fakulta

DOI

https://doi.org/10.1021/acs.analchem.2c05670

UT WoS

000939526700001

EID Scopus

2-s2.0-85149135669

Klíčová slova anglicky

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

Štítky

14313050, CF NANO, H24, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 2. 11. 2024 15:27, 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

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://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 = {https://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://doi.org/10.1021/acs.analchem.2c05670.

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