Background-free cell surface glycan analysis using persistent luminescence nanoparticle as an optical probe

• Published in: Analytical biochemistry 2020-07, Vol.601, p.113780, Article 113780
• Main Authors: He, Na, Jiang, Ying, Lei, Lingli, Liu, Yingshuai
• Format: Article
• Language: English
• Subjects: Background-free analysis; Cell surface glycans; Cells, Cultured; Humans; Lectins; Luminescence; Luminescent Agents - chemistry; Nanoparticles - chemistry; Optical Imaging; Persistent luminescence nanoparticles; Polysaccharides - analysis
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2020.113780

In this work, we report a novel cell surface glycan analysis method based on persistent luminescence nanoparticle (PLNP) ZnGa2O4: Cr3+ (ZGC) as an optical probe. ZGC was first silanized by (3-Aminopropyl) triethoxysilane (APTES), followed by PEGylation with NHS-P EG-Biotin, which not only introduces biotin, but significantly improves the dispersibility and stability of the nanoparticles. Neutral-avidin was then coupled on ZGC surface through the specific biotin-avidin interaction, producing a ZGC-PEG-avidin nanoprobe. As for cell surface glycan detection, different surface glycans are recognized with their corresponding biotinylated lectins, which are then traced by ZGC-PEG-avidin. The persistent luminescence signal is recorded by a microtiter plate reader in time-resolved fluorescence mode. Glycans expression profiling on prostate cancer cell DU145 and normal prostate cell RWPE-1 was analyzed by the proposed detection platform. Similar results were observed from the conventional horseradish peroxidase (HRP)-catalyzed absorbent assay and confocal microscope-based fluorescence imaging, demonstrating the applicability of the proposed platform. The approach based on the long afterglow property of ZGC efficiently eliminates the background noise from cells and substrate, resulting in the best signal-to-noise ratio and high detection sensitivity. •ZGC-PEG-Biotin with good dispersibility and persistent luminescence is synthesized by PEGylation.•A background-free analytical platform is developed with ZGC as an optical nanoprobe.•Glycan expression profiling on cancer and normal cell surface was analyzed by the proposed platform.