k 2021

Near-infrared polymethine fluorophores for targeted biomarker detection

PASTUCHA, Matěj, Martin POTRUSIL, Zdeněk FARKA and Peter ŠEBEJ

Basic information

Original name

Near-infrared polymethine fluorophores for targeted biomarker detection

Authors

Edition

XX. Interdisciplinary meeting of young life scientists, 2021

Other information

Language

English

Type of outcome

Prezentace na konferencích

Field of Study

10406 Analytical chemistry

Country of publisher

Czech Republic

Confidentiality degree

není předmětem státního či obchodního tajemství

Organization unit

Faculty of Science
Změněno: 21/5/2021 18:18, doc. Mgr. Zdeněk Farka, Ph.D.

Abstract

V originále

Fluorescent dyes are widely used in life-sciences and medicine. They are frequently employed as imaging agents, alone or in connection with antibodies in immunofluorescence experiments. Biological tissues absorb visible light; therefore, a tissue-transparent window (TTW) in the near-infrared (NIR) region is preferred for imaging purposes, e.g. in the fluorescence image-guided surgery. Polymethine dyes are great candidates for applications in the TTW, and their properties can be tuned by changes in the chromophore length and substituents. We chose the indocyanine green (ICG) as an initial model (the only NIR emitting dye certified by FDA for use in clinical medicine). It is usually used as a contrast agent with limited selectivity, but when conjugated to an antibody, it can also be employed for specific visualization of target biomarkers. However, its conjugation is not without pitfalls. The dye inflicted partial aggregation of the antibody during conjugation. Furthermore, we observed a high level of nonspecific binding in a model immunocytochemical labeling of HER2 biomarker on breast cancer cell lines. This was probably caused by the high tendency of the dye to bind to proteins. To overcome this limitation, we conjugated a heptamethine with similar spectral properties, Cyanine 7 (Cy7), to the antibody via Cu-catalyzed click chemistry. The conjugate provided a much higher specificity of labeling, making it a superior NIR dye for immunofluorescence experiments in the TTW.

Links

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