A Permeabilization Workflow To Enable Specific Multiplexed Profiling Using SERS Nanoparticles.

Area of Science:

• Biomedical Engineering
• Nanotechnology
• Cancer Diagnostics

Background:

• Surface-enhanced Raman scattering nanoparticles (SERS NPs) show promise for cellular targeting and biomarker detection.
• Current limitations include poor membrane permeability and endosomal trapping, hindering intracellular applications.

Purpose of the Study:

• To develop and optimize a SERS NP-based strategy for robust intracellular targeting of breast cancer biomarkers.
• To demonstrate multiplexed detection of both extracellular (HER2) and intracellular (ER, PR) targets.

Main Methods:

• Conjugation of SERS NPs with antibodies against HER2, ER, and PR.
• Evaluation of permeabilization agents (Tween 20, Triton X-100, methanol) for intracellular delivery.
• Assessment of SERS NP binding specificity and cell viability using flow cytometry, fluorescence, and Raman imaging.

Main Results:

• Triton X-100 was identified as the optimal permeabilization agent, allowing SERS NP entry while maintaining cell viability.
• High specificity (>100-fold) was observed for HER2-overexpressing cells.
• Successful intracellular targeting and detection of ER and PR were achieved, alongside multiplexed detection of HER2, ER, and PR.

Conclusions:

• The optimized SERS NP strategy enables efficient intracellular biomarker detection in breast cancer cells.
• This platform offers potential for enhanced, multiplexed diagnostics, including applications in liquid biopsies.
• The findings advance the use of SERS NPs for improved breast cancer diagnosis and personalized medicine.