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Updated: Jul 26, 2025

Colorectal Cancer Cell Surface Protein Profiling Using an Antibody Microarray and Fluorescence Multiplexing
Published on: September 25, 2011
SERS characterization of colorectal cancer cell surface markers upon anti-EGFR treatment
Nana Lyu1, Bernadette Pedersen2, Elena Shklovskaya2
1ARC Center of Excellence for Nanoscale BioPhotonics and School of Natural Sciences, Faculty of Science and Engineering Macquarie University Sydney New South Wales Australia.
Abstract:
Colorectal cancer (CRC) is the third most diagnosed and the second lethal cancer worldwide. Approximately 30-50% of CRC are driven by mutations in the KRAS oncogene, which is a strong negative predictor for response to anti-epidermal growth factor receptor (anti-EGFR) therapy. Examining the phenotype of KRAS mutant and wild-type (WT) CRC cells in response to anti-EGFR treatment may provide significant insights into drug response and resistance. Herein, surface-enhanced Raman spectroscopy (SERS) assay was applied to phenotype four cell surface proteins (EpCAM, EGFR, HER2, HER3) in KRAS mutant (SW480) and WT (SW48) cells over a 24-day time course of anti-EGFR treatment with cetuximab. Cell phenotypes were obtained using Raman reporter-coated and antibody-conjugated gold nanoparticles (SERS nanotags), where a characteristic Raman spectrum was generated upon single laser excitation, reflecting the presence of the targeted surface marker proteins. Compared to the KRAS mutant cells, KRAS WT cells were more sensitive to anti-EGFR treatment and displayed a significant decrease in HER2 and HER3 expression. The SERS results were validated with flow cytometry, confirming the SERS assay is promising as an alternative method for multiplexed characterization of cell surface biomarkers using a single laser excitation system.
Insights
Surface-enhanced Raman spectroscopy (SERS) effectively characterized colorectal cancer (CRC) cells. KRAS wild-type CRC cells showed increased sensitivity to anti-EGFR therapy, with decreased HER2 and HER3 expression.
Area of Science:
- Oncology
- Biotechnology
- Spectroscopy
Background:
- Colorectal cancer (CRC) is a leading cause of cancer death globally.
- KRAS mutations in CRC predict poor response to anti-EGFR therapies.
- Understanding KRAS-driven phenotypes is crucial for improving CRC treatment.
Purpose of the Study:
- To phenotype KRAS mutant and wild-type (WT) CRC cells under anti-EGFR treatment.
- To investigate cell surface protein expression changes during treatment.
- To evaluate Surface-Enhanced Raman Spectroscopy (SERS) as a method for biomarker characterization.
Main Methods:
- A SERS assay using antibody-conjugated gold nanoparticles (nanotags) was developed.
- Four cell surface proteins (EpCAM, EGFR, HER2, HER3) were phenotyped.
- KRAS mutant (SW480) and WT (SW48) CRC cells were treated with cetuximab (anti-EGFR) over 24 days.
Main Results:
- KRAS WT cells exhibited higher sensitivity to anti-EGFR treatment compared to KRAS mutant cells.
- A significant decrease in HER2 and HER3 expression was observed in KRAS WT cells.
- SERS assay results were validated by flow cytometry.
Conclusions:
- SERS is a promising method for multiplexed cell surface biomarker analysis.
- Phenotypic differences in KRAS WT and mutant CRC cells influence anti-EGFR therapy response.
- SERS offers a potential alternative for characterizing cell surface markers with a single laser system.

