Isolating epidermal growth factor receptor overexpressing carcinoma cells from human whole blood by bio‐ferrography. Issue 2 (10th December 2014)

• Record Type: Journal Article
• Authors: Levi, Ofer; Shapira, Assaf; Tal, Baruch; Benhar, Itai; Eliaz, Noam
• Is Part Of: Cytometry. Volume 88:Issue 2(2015)
• Page Start: 136
• Page End: 144
• Publication Date: 2014-12-10
• Subjects: A431 cells -- Bio‐Ferrography -- EGFR -- human whole blood -- monoclonal antibody -- immunomagnetic separation; Flow cytometry -- Diagnostic use -- Periodicals; Cytodiagnosis -- Periodicals; 616.07582
• DOI: 10.1002/cyto.b.21212
• Languages: English
• ISSNs: 1552-4949

Abstract

Abstract : Background: The epidermal growth factor receptor (EGFR) is overexpressed in carcinoma. In some cases, including in colorectal cancer, it is used as a therapeutic target. Bio‐Ferrography is a nondestructive method for isolating magnetized cells and tissues from a fluid onto a glass slide based on their interaction with an external, strong, and focused magnetic field. Methods: Here, we implement Bio‐Ferrography to separate EGFR‐positive cancer cells from EGFR‐negative noncancer cells, mixed at a ratio of 1 to 1 × 10 6, from either phosphate‐buffered saline or human whole blood. Incubation of the cells with an anti‐EGFR antibody and magnetic microbeads coupled to a secondary antibody was used to magnetize the target cells prior to the ferrographic analysis. Results: A procedure was developed for "a proof of concept" isolation. Recovery values as high as 78% for 1 mL phosphate‐buffered saline, and 53% for 1 mL human whole blood, with a limit‐of‐detection of 30 and 100 target cells, respectively, were achieved. Conclusions: These capture efficiency values are considered significant and, therefore, warrant further study on isolation of real circulating tumor cells from blood samples of patients, aiming at early diagnosis of EGFR‐overexpressing tumor types. © 2014 International Clinical Cytometry Society