Combining rVAR2 and Anti-EpCAM to Increase the Capture Efficiency of Non-Small-Cell Lung Cancer Cell Lines in the Flow Enrichment Target Capture Halbach (FETCH) Magnetic Separation System

Int J Mol Sci. 2024 Sep 11;25(18):9816. doi: 10.3390/ijms25189816.

Abstract

Circulating tumor cells (CTCs) are detected in approximately 30% of metastatic non-small-cell lung cancer (NSCLC) cases using the CellSearch system, which relies on EpCAM immunomagnetic enrichment and Cytokeratin detection. This study evaluated the effectiveness of immunomagnetic enrichment targeting oncofetal chondroitin sulfate (ofCS) using recombinant VAR2CSA proteins (rVAR2) to improve the recovery of different NSCLC cell lines spiked into lysed blood samples. Four NSCLC cell lines-NCI-H1563, A549, NCI-H1792, and NCI-H661-were used to assess capture efficiency. The results demonstrated that the combined use of anti-EpCAM antibody and rVAR2 significantly enhanced the capture efficiency to an average of 88.2% compared with 40.6% when using only anti-EpCAM and 56.6% when using only rVAR2. These findings suggest that a dual-marker approach using anti-EpCAM and rVAR2 can provide a more robust and sensitive method for CTC enrichment in NSCLC, potentially leading to better diagnostic and prognostic outcomes.

Keywords: EpCAM; circulating tumor cells (CTCs); immunomagnetic enrichment; non-small-cell lung cancer (NSCLC); rVAR2.

MeSH terms

  • A549 Cells
  • Antigens, Neoplasm / immunology
  • Antigens, Neoplasm / metabolism
  • Antigens, Protozoan
  • Biomarkers, Tumor
  • Carcinoma, Non-Small-Cell Lung* / metabolism
  • Carcinoma, Non-Small-Cell Lung* / pathology
  • Cell Line, Tumor
  • Chondroitin Sulfates / metabolism
  • Epithelial Cell Adhesion Molecule* / metabolism
  • Humans
  • Immunomagnetic Separation / methods
  • Lung Neoplasms* / immunology
  • Lung Neoplasms* / metabolism
  • Lung Neoplasms* / pathology
  • Neoplastic Cells, Circulating* / metabolism
  • Neoplastic Cells, Circulating* / pathology
  • Recombinant Proteins

Substances

  • Epithelial Cell Adhesion Molecule
  • EPCAM protein, human
  • Biomarkers, Tumor
  • VAR2CSA protein, Plasmodium falciparum
  • Recombinant Proteins
  • Antigens, Neoplasm
  • Chondroitin Sulfates
  • Antigens, Protozoan