Negative Charge-Carrying Glycans Attached to Exosomes as Novel Liquid Biopsy Marker
Abstract
:1. Introduction
2. Materials and Methods
2.1. Chemicals
2.2. Cell Culture Cultivation
2.3. Exosome Collection and Isolation
2.4. Nanoparticle-Tracking Analysis (NTA)
2.5. Dynamic Light-Scattering (DLS)
2.6. Microscopic Analysis (SEM and AFM)
2.7. Surface Plasmon Resonance (SPR)
2.8. Electrochemical Measurements
3. Results
3.1. NTA Characterisation
3.2. DLS and ζ-Potential Characterisation
3.3. Microscopic Characterisation of Isolated Exosomes
3.4. SPR Glycoprofiling
3.4.1. Antibody Selection
3.4.2. Lectin-Based In-Situ Glycoprofiling
3.4.3. SPR Sandwich Configuration
3.5. Electrochemical Measurements
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Lectin/Agglutinin (Abbreviation) | Specificity | SU/GBS | Carcinoma Exosomes | Control Exosomes |
---|---|---|---|---|
Aleuria aurantia (AAL) | fucose, arabinose | 2 subunits/5 fucose BS | − | − |
Galanthus nivalis (GNL) | mannose | 4 subunits/12 mannose BS | − | − |
Maackia amurensis (MAAII) | α-2,3 linked Sia | 2 subunits | + | + |
Phaseolus vulgaris (PHA-E) | galactose, bisecting GlcNac, biantennary N-glycans | 4 E subunits | − | + |
Phaseolus vulgaris (PHA-L) | branched N-glycans | 4 L type subunits | − | + |
Pisum sativum (PSA) | mannose, glucose | 4 subunits | − | − |
Sambucus nigra (SNA) | α-2,6 linked Sia | 2 subunits | + | + |
Wisteria floribunda (WFL) | N-Acetylgalactosamine | 4 subunits | − | − |
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Kosutova, N.; Lorencova, L.; Hires, M.; Jane, E.; Orovcik, L.; Kollar, J.; Kozics, K.; Gabelova, A.; Ukraintsev, E.; Rezek, B.; et al. Negative Charge-Carrying Glycans Attached to Exosomes as Novel Liquid Biopsy Marker. Sensors 2024, 24, 1128. https://doi.org/10.3390/s24041128
Kosutova N, Lorencova L, Hires M, Jane E, Orovcik L, Kollar J, Kozics K, Gabelova A, Ukraintsev E, Rezek B, et al. Negative Charge-Carrying Glycans Attached to Exosomes as Novel Liquid Biopsy Marker. Sensors. 2024; 24(4):1128. https://doi.org/10.3390/s24041128
Chicago/Turabian StyleKosutova, Natalia, Lenka Lorencova, Michal Hires, Eduard Jane, Lubomir Orovcik, Jozef Kollar, Katarina Kozics, Alena Gabelova, Egor Ukraintsev, Bohuslav Rezek, and et al. 2024. "Negative Charge-Carrying Glycans Attached to Exosomes as Novel Liquid Biopsy Marker" Sensors 24, no. 4: 1128. https://doi.org/10.3390/s24041128