Near-infrared luminescent labels for immunochemical assays and
imaging

PASTUCHA, Matěj, Zdeněk FARKA, Peter ŠEBEJ, Antonín HLAVÁČEK and Petr SKLÁDAL. Near-infrared luminescent labels for immunochemical assays and imaging. In 72. sjezd českých a slovenských chemických společností. 2020. ISSN 2336-7202.

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TY  - CONF
ID  - 1678220
AU  - Pastucha, Matěj - Farka, Zdeněk - Šebej, Peter - Hlaváček, Antonín - Skládal, Petr
PY  - 2020
TI  - Near-infrared luminescent labels for immunochemical assays and imaging
UR  - https://sjezd72.csch.cz/wp-content/uploads/2020/09/Sjezd-Praha-2020-tisk.pdf
N2  - Fluorescent dyes are widely applied in life sciences and medicine as imaging agents, either alone or conjugated with antibodies or other recognition elements to allow targeting. However, when working with tissues, absorption of visible light and autofluorescence is limiting the use of traditional fluorophores. Therefore, probes working in the far-red and near-infrared (NIR) tissue transparent window are preferred. Among small molecules, polymethine dyes are suitable candidates. Their properties can be tuned by changes in the polymethine chain length and substituents. We chose the indocyanine green (ICG) as an initial model. It is the only NIR fluorophore certified by the FDA for use in clinical medicine. Traditionally, it is used unconjugated as an imaging agent in fluorescence image-guided surgery. Upon coupling with a primary antibody, we have adopted it for specific fluorescent detection of the cancer biomarker CA 19-9. Nanoparticles are an emerging alternative to conventional fluorophores. Despite several limitations, mainly related to their size and toxicity, the photon-upconversion nanoparticles (UCNPs) are particularly attractive. They can convert absorbed NIR light to shorter wavelengths. Thanks to this anti-Stokes emission, the autofluorescence from the sample is eliminated. In comparison to small fluorophores, the surface of the NPs plays a more critical role. Surface modification becomes a necessity but also an opportunity to tune the nanoparticle stability, anti-fouling properties, and biocompatibility. We have applied the BSA- and streptavidin-modified UCNPs as labels in a microtiter plate-based immunoassay for the detection of bacterial honeybee pathogens. Furthermore, surface modification by PEG-neridronate made the UCNPs useful in immunocytochemical staining, which opens the way for their use in more complex biological systems.
ER  -

Basic information
Original name	Near-infrared luminescent labels for immunochemical assays and imaging
Authors	PASTUCHA, Matěj, Zdeněk FARKA, Peter ŠEBEJ, Antonín HLAVÁČEK and Petr SKLÁDAL.
Edition	72. sjezd českých a slovenských chemických společností, 2020.

Other information
Original language	English
Type of outcome	Conference abstract
Field of Study	10406 Analytical chemistry
Country of publisher	Czech Republic
Confidentiality degree	is not subject to a state or trade secret
WWW	URL
Organization unit	Faculty of Science
ISSN	2336-7202
Changed by	Changed by: doc. Mgr. Zdeněk Farka, Ph.D., učo 357740. Changed: 10/9/2020 22:46.

Abstract

Fluorescent dyes are widely applied in life sciences and medicine as imaging agents, either alone or conjugated with antibodies or other recognition elements to allow targeting. However, when working with tissues, absorption of visible light and autofluorescence is limiting the use of traditional fluorophores. Therefore, probes working in the far-red and near-infrared (NIR) tissue transparent window are preferred. Among small molecules, polymethine dyes are suitable candidates. Their properties can be tuned by changes in the polymethine chain length and substituents. We chose the indocyanine green (ICG) as an initial model. It is the only NIR fluorophore certified by the FDA for use in clinical medicine. Traditionally, it is used unconjugated as an imaging agent in fluorescence image-guided surgery. Upon coupling with a primary antibody, we have adopted it for specific fluorescent detection of the cancer biomarker CA 19-9. Nanoparticles are an emerging alternative to conventional fluorophores. Despite several limitations, mainly related to their size and toxicity, the photon-upconversion nanoparticles (UCNPs) are particularly attractive. They can convert absorbed NIR light to shorter wavelengths. Thanks to this anti-Stokes emission, the autofluorescence from the sample is eliminated. In comparison to small fluorophores, the surface of the NPs plays a more critical role. Surface modification becomes a necessity but also an opportunity to tune the nanoparticle stability, anti-fouling properties, and biocompatibility. We have applied the BSA- and streptavidin-modified UCNPs as labels in a microtiter plate-based immunoassay for the detection of bacterial honeybee pathogens. Furthermore, surface modification by PEG-neridronate made the UCNPs useful in immunocytochemical staining, which opens the way for their use in more complex biological systems.

Links
GJ20-30004Y, research and development project	Name: Nové strukturní motivy pro rozšíření možností lineárních polymethinových barviv
GJ20-30004Y, research and development project	Investor: Czech Science Foundation
LQ1601, research and development project	Name: CEITEC 2020 (Acronym: CEITEC2020)
LQ1601, research and development project	Investor: Ministry of Education, Youth and Sports of the CR
LTAB19011, research and development project	Name: Luminiscenční imunostanovení jako citlivý nástroj pro diagnostiku nemocí včel
LTAB19011, research and development project	Investor: Ministry of Education, Youth and Sports of the CR, Sensitive bioanalytical tools for the surveillance of honey bee diseases, INTER-ACTION

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Near-infrared luminescent labels for immunochemical assays and imaging

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