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Nanopipette-Based SERS Aptasensor for Subcellular Localization of Cancer Biomarker in Single Cells
Sumaira Hanif1, Hai-Ling Liu1, Saud Asif Ahmed1
1State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, and ‡State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University , Nanjing 210023, China.
This study introduces a novel nanopipette technique for precise cancer cell analysis. It enhances specificity in detecting Nucleolin (NCL) protein, crucial for cancer diagnosis and treatment, by minimizing cellular matrix effects.
Area of Science:
- Biomedical Engineering
- Analytical Chemistry
- Molecular Biology
Background:
- Single cell analysis is vital for understanding cellular heterogeneity and cancer progression.
- Nucleolin (NCL) is overexpressed in cancer cells, influencing proliferation and treatment response.
- Existing methods for cancer biomarker detection lack specificity due to subcellular matrix effects.
Purpose of the Study:
- To develop a novel technique for enhanced molecular and spectral specificity in cancer diagnosis.
- To improve the detection of Nucleolin (NCL) protein at the subcellular level.
- To enable precise single-cell analysis for cancer diagnosis and treatment.
Main Methods:
- Utilized aptamer-functionalized gold nanopipettes for target capture.
- Employed p-mercaptobenzonitrile (MBN) and DNA-modified Ag nanoparticles as Surface-Enhanced Raman Scattering (SERS) reporters.
- Developed a nanoprobe system that loses SERS signal upon specific NCL capture.
Main Results:
- Achieved specific extraction of NCL protein from various cell lines (MCF-7A, HeLa, MCF-10A).
- Demonstrated high affinity for NCL with a dissociation constant (Kd) of 36 nM.
- Showcased 1000-fold higher specificity against competing proteins and negligible matrix effects.
Conclusions:
- The developed aptamer-based SERS nanopipette offers high specificity and sensitivity for NCL detection.
- Successfully mapped subcellular localization and spatial distribution of NCL in cancer cells.
- This technique provides a new perspective for single-cell cancer diagnosis and targeted therapy.
