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Background-free cell surface glycan analysis using persistent luminescence nanoparticle as an optical probe.
Na He1, Ying Jiang1, Lingli Lei1
1Institute for Clean Energy and Advanced Materials, School of Materials and Energy, Southwest University, Chongqing, 400715, PR China.
Analytical Biochemistry
|May 30, 2020
Summary
We developed a new method for analyzing cell surface glycans using persistent luminescence nanoparticles (PLNPs). This sensitive optical probe offers high detection sensitivity and an excellent signal-to-noise ratio for glycan expression profiling.
Area of Science:
- Biotechnology
- Nanotechnology
- Analytical Chemistry
Background:
- Cell surface glycans play crucial roles in biological processes.
- Accurate analysis of glycan expression is vital for understanding diseases like cancer.
- Existing methods for glycan analysis can be limited by background noise and sensitivity.
Purpose of the Study:
- To develop a novel, highly sensitive optical probe for cell surface glycan analysis.
- To utilize persistent luminescence nanoparticles (PLNPs) for improved detection sensitivity.
- To validate the proposed method for glycan expression profiling in cancer cells.
Main Methods:
- Functionalization of persistent luminescence nanoparticle (PLNP) ZnGa2O4: Cr3+ (ZGC) with APTES and PEG-Biotin.
- Coupling of neutral-avidin to create a ZGC-PEG-avidin nanoprobe.
- Detection of cell surface glycans using biotinylated lectins and the ZGC-PEG-avidin nanoprobe.
- Measurement of persistent luminescence signals using a microtiter plate reader in time-resolved fluorescence mode.
- Analysis of glycan expression on DU145 prostate cancer cells and RWPE-1 normal prostate cells.
Main Results:
- The ZGC-PEG-avidin nanoprobe demonstrated effective detection of cell surface glycans.
- Glycan expression profiling on prostate cancer cells (DU145) and normal cells (RWPE-1) was successfully performed.
- Results were consistent with conventional methods like HRP-catalyzed assays and fluorescence imaging.
- The persistent luminescence approach significantly reduced background noise, leading to superior signal-to-noise ratio.
Conclusions:
- The developed PLNP-based method offers a sensitive and reliable platform for cell surface glycan analysis.
- The long afterglow property of ZGC nanoparticles enhances detection sensitivity and minimizes background interference.
- This novel approach holds promise for accurate glycan expression profiling in various biological and clinical applications.
