Optimization of EGFR high positive cell isolation procedure by design of experiments methodology

Cytometry B Clin Cytom. 2015 Sep-Oct;88(5):338-47. doi: 10.1002/cyto.b.21246. Epub 2015 Apr 18.

Abstract

Background: Circulating tumor cells (CTCs) in blood circulation may play a role in monitoring and even in early detection of metastasis patients. Due to the limited presence of CTCs in blood circulation, viable CTCs isolation technology must supply a very high recovery rate.

Methods: Here, we implement design of experiments (DOE) methodology in order to optimize the Bio-Ferrography (BF) immunomagnetic isolation (IMI) procedure for the EGFR high positive CTCs application. All consequent DOE phases such as screening design, optimization experiments and validation experiments were used.

Results: A significant recovery rate of more than 95% was achieved while isolating 100 EGFR high positive CTCs from 1 mL human whole blood.

Conclusions: The recovery achievement in this research positions BF technology as one of the most efficient IMI technologies, which is ready to be challenged with patients' blood samples.

Keywords: EGFR; bio-ferrography; carcinoma; design of experiments; flow cytometry; immunomagnetic isolation.

MeSH terms

  • Biomarkers, Tumor / analysis*
  • Cell Line, Tumor
  • Equipment Design
  • ErbB Receptors / analysis*
  • Flow Cytometry / instrumentation
  • Flow Cytometry / methods*
  • Humans
  • Immunomagnetic Separation / instrumentation
  • Immunomagnetic Separation / methods*
  • Neoplastic Cells, Circulating / chemistry*

Substances

  • Biomarkers, Tumor
  • EGFR protein, human
  • ErbB Receptors