Synthesis and application of BODIPY-based fluorescent labeling
tag for oligosaccharide and N-linked glycan analysis by
high-performance liquid chromatography with fluorescence
detection

J 2024

Synthesis and application of BODIPY-based fluorescent labeling tag for oligosaccharide and N-linked glycan analysis by high-performance liquid chromatography with fluorescence detection

SMOLKOVÁ, Denisa; Michal GREGUŠ; Hubert VESELÝ; Richard CMELIK; Hana PÍŽOVÁ et al.

Základní údaje

Originální název

Synthesis and application of BODIPY-based fluorescent labeling tag for oligosaccharide and N-linked glycan analysis by high-performance liquid chromatography with fluorescence detection

Autoři

SMOLKOVÁ, Denisa; Michal GREGUŠ; Hubert VESELÝ; Richard CMELIK; Hana PÍŽOVÁ; Pavel BOBÁĽ a Jana LAVICKA

Vydání

Analytica Chimica Acta, Amsterdam, Elsevier Science publishers, 2024, 0003-2670

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10400 1.4 Chemical sciences

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 6.000

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14160/24:00135614

Organizační jednotka

Farmaceutická fakulta

Klíčová slova anglicky

Oligosaccharides; Glycans; Labeling; BODIPY; Liquid chromatography; Fluorescence

Štítky

14313010, rivok, ÚChL

Příznaky

Recenzováno

Změněno: 27. 8. 2025 08:23, Mgr. Marie Novosadová Šípková, DiS.

Anotace

V originále

Background: Glycosylation analysis is still challenging, not only because of the extreme structure complexity and conjugation diversity of glycans but also because of instrumental aspects such as the sensitivity limits of analyses. Therefore, glycan analysis by chromatographic methods is very often combined with fluorescence detection in addition to MS. The majority of fluorescent labeling employed before LC separation is based on 2-aminobenzamide, which has several disadvantages such as low labeling yield, poor fluorescence properties, and MS ionization efficiency. Therefore, even after several decades of development of new labels, there is still a need for new labeling tags with improved characteristics. Results: We present the application of a newly synthesized fluorescent label designed for oligosaccharide and glycan analysis by high-performance liquid chromatography with fluorescence detection (HPLC/FLD). The novel hydrazide derivative of dipyrrometheneboron difluoride (BODIPY) was synthesized from 2,4-dimethylpyrrole, methyl succinyl chloride, and boron trifluoride etherate followed by a reaction with hydrazine. The synthesized label was characterized by several analytical methods including NMR, UV/Vis and fluorescence spectroscopy, and mass spectrometry. The labeling reaction via hydrazone formation chemistry was optimized by labeling of maltooligosaccharide standards. The analysis of maltohexaose labeled by BODIPY-hydrazide followed by HPLC/ FLD analysis provided the limit of detection in the low tens of femtomole. The presented method based on fluorescence detection is at least 30 times more sensitive than the standard approach employing labeling by 2aminobenzamide. In addition, the labeling method by BODIPY-hydrazide was used for N -linked glycan profiling of several glycoproteins (ribonuclease B, immunoglobulin G) by RP-HPLC/FLD as well as HILIC/FLD analysis. Significance: This work represents the design, synthesis, and application of a new fluorescent label based on the BODIPY core and hydrazone formation chemistry for oligosaccharide and glycan analysis by HPLC/FLD. The proposed approach significantly improved the oligosaccharide and glycan analysis in comparison to the commonly used procedure employing 2-aminobenzamide.

Návaznosti

GA22-00236S, projekt VaV

Název: Vývoj nových fluorescenčních značek pro elektroforetické analýzy glykoproteinů asociovaných s rakovinou prsu (Akronym: 22-00236S)

Investor: Grantová agentura ČR, Vývoj nových fluorescenčních značek pro elektroforetické analýzy glykoproteinů asociovaných s rakovinou prsu

MUNI/A/1232/2021, interní kód MU

Název: Design a syntéza nových fluoroforů pro detekci biomolekul

Investor: Masarykova univerzita, Design a syntéza nových fluoroforů pro detekci biomolekul

Citovat

SMOLKOVÁ, Denisa; Michal GREGUŠ; Hubert VESELÝ; Richard CMELIK; Hana PÍŽOVÁ; Pavel BOBÁĽ a Jana LAVICKA. Synthesis and application of BODIPY-based fluorescent labeling tag for oligosaccharide and N-linked glycan analysis by high-performance liquid chromatography with fluorescence detection. Analytica Chimica Acta. Amsterdam: Elsevier Science publishers, 2024, roč. 1285, č. 342032, s. 1-8. ISSN 0003-2670. Dostupné z: https://doi.org/10.1016/j.aca.2023.342032.

@article{2380465,
   author = {Smolková, Denisa and Greguš, Michal and Veselý, Hubert and Cmelik, Richard and Pížová, Hana and Bobáľ, Pavel and Lavicka, Jana},
   article_location = {Amsterdam},
   article_number = {342032},
   doi = {https://doi.org/10.1016/j.aca.2023.342032},
   keywords = {Oligosaccharides; Glycans; Labeling; BODIPY; Liquid chromatography; Fluorescence},
   language = {eng},
   issn = {0003-2670},
   journal = {Analytica Chimica Acta},
   title = {Synthesis and application of BODIPY-based fluorescent labeling tag for oligosaccharide and N-linked glycan analysis by high-performance liquid chromatography with fluorescence detection},
   url = {https://pdf.sciencedirectassets.com/271374/1-s2.0-S0003267023X00464/1-s2.0-S0003267023012539/main.pdf?X-Amz-Security-Token=IQoJb3JpZ2luX2VjEFkaCXVzLWVhc3QtMSJHMEUCIQCfgifaC8tRpXFB%2F4tVDJ4386lwIOmxH38Q0ElbRrTFTQIgaEba7F3qlS8%2ByHAwZVa4RfNShZVFB%2B12U%2BX},
   volume = {1285},
   year = {2024}
}

TY  - JOUR
ID  - 2380465
AU  - Smolková, Denisa - Greguš, Michal - Veselý, Hubert - Cmelik, Richard - Pížová, Hana - Bobáľ, Pavel - Lavicka, Jana
PY  - 2024
TI  - Synthesis and application of BODIPY-based fluorescent labeling tag for oligosaccharide and N-linked glycan analysis by high-performance liquid chromatography with fluorescence detection
JF  - Analytica Chimica Acta
VL  - 1285
IS  - 342032
SP  - 1-8
EP  - 1-8
PB  - Elsevier Science publishers
SN  - 00032670
KW  - Oligosaccharides
KW  - Glycans
KW  - Labeling
KW  - BODIPY
KW  - Liquid chromatography
KW  - Fluorescence
UR  - https://pdf.sciencedirectassets.com/271374/1-s2.0-S0003267023X00464/1-s2.0-S0003267023012539/main.pdf?X-Amz-Security-Token=IQoJb3JpZ2luX2VjEFkaCXVzLWVhc3QtMSJHMEUCIQCfgifaC8tRpXFB%2F4tVDJ4386lwIOmxH38Q0ElbRrTFTQIgaEba7F3qlS8%2ByHAwZVa4RfNShZVFB%2B12U%2BX
N2  - Background: Glycosylation analysis is still challenging, not only because of the extreme structure complexity and conjugation diversity of glycans but also because of instrumental aspects such as the sensitivity limits of analyses. Therefore, glycan analysis by chromatographic methods is very often combined with fluorescence detection in addition to MS. The majority of fluorescent labeling employed before LC separation is based on 2-aminobenzamide, which has several disadvantages such as low labeling yield, poor fluorescence properties, and MS ionization efficiency. Therefore, even after several decades of development of new labels, there is still a need for new labeling tags with improved characteristics. Results: We present the application of a newly synthesized fluorescent label designed for oligosaccharide and glycan analysis by high-performance liquid chromatography with fluorescence detection (HPLC/FLD). The novel hydrazide derivative of dipyrrometheneboron difluoride (BODIPY) was synthesized from 2,4-dimethylpyrrole, methyl succinyl chloride, and boron trifluoride etherate followed by a reaction with hydrazine. The synthesized label was characterized by several analytical methods including NMR, UV/Vis and fluorescence spectroscopy, and mass spectrometry. The labeling reaction via hydrazone formation chemistry was optimized by labeling of maltooligosaccharide standards. The analysis of maltohexaose labeled by BODIPY-hydrazide followed by HPLC/ FLD analysis provided the limit of detection in the low tens of femtomole. The presented method based on fluorescence detection is at least 30 times more sensitive than the standard approach employing labeling by 2aminobenzamide. In addition, the labeling method by BODIPY-hydrazide was used for N -linked glycan profiling of several glycoproteins (ribonuclease B, immunoglobulin G) by RP-HPLC/FLD as well as HILIC/FLD analysis. Significance: This work represents the design, synthesis, and application of a new fluorescent label based on the BODIPY core and hydrazone formation chemistry for oligosaccharide and glycan analysis by HPLC/FLD. The proposed approach significantly improved the oligosaccharide and glycan analysis in comparison to the commonly used procedure employing 2-aminobenzamide.
ER  -

SMOLKOVÁ, Denisa; Michal GREGUŠ; Hubert VESELÝ; Richard CMELIK; Hana PÍŽOVÁ; Pavel BOBÁĽ a Jana LAVICKA. Synthesis and application of BODIPY-based fluorescent labeling tag for oligosaccharide and N-linked glycan analysis by high-performance liquid chromatography with fluorescence detection. \textit{Analytica Chimica Acta}. Amsterdam: Elsevier Science publishers, 2024, roč.~1285, č.~342032, s.~1-8. ISSN~0003-2670. Dostupné z: https://doi.org/10.1016/j.aca.2023.342032.

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