title: A mesofluidic multiplex immunosensor for detection of circulating cytokeratin-positive cells in the blood of breast cancer patients.
abstract: We have recently reported the analytical performance of an immunosensor comprising one mm-scale parallel plate laminar flow chamber and applied to capture MCF7 breast cancer cells (Ehrhart et al., Biosens. Bioelectr. 24, 467, 2008). Herein we present a new multiplex immunosensor embodying four parallel plate laminar flow chambers that fit onto a standard, functionalized, microscopy glass slide. The four surfaces are coated with long alkyl chain spacers of 21-aminohenicosyl trichlorosilane (AHTS) and then grafted with a monoclonal anti-human epithelial cell adhesion molecule (EpCAM) antibody specific of target cells to immobilize. We first demonstrate a significantly (P < 0.01) improved capacity of each of the four flow chambers of the multiplex immunosensor to capture MCF7 cells compared to the previous single chamber device. Second, in addition to an increase of cell immobilization, the multiplex device offers a versatile tool easily grafted with various purified antibodies onto the four surfaces. Third, we obtained high cell capture rate and efficiency of various numbers of MCF7 cells spiked in buffer containing an equal number of background leukocytes. And fourth, we demonstrate isolation efficiency of circulating tumor cells (CTCs) from peripheral blood drawn from a small cohort of patients with localized or metastatic breast cancer. This new multiplex immunosensor could be tested for its potential to capture different subpopulations of CTCs.
authors.affiliation: CyToCap, ENS Cachan-Institut d'Alembert, 61 Avenue du Président Wilson, 94235 Cachan Cedex, France.
authors.forename: François
authors.initials: F
authors.lastname: Breton
authors.forename: Bernard
authors.initials: B
authors.lastname: Bennetau
authors.forename: Rosette
authors.initials: R
authors.lastname: Lidereau
authors.forename: Laurent
authors.initials: L
authors.lastname: Thomas
authors.forename: Gilles
authors.initials: G
authors.lastname: Regnier
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Ehrhart
authors.forename: Patrick
authors.initials: P
authors.lastname: Tauc
authors.forename: Phuong-Lan
authors.initials: PL
authors.lastname: Tran
mesh_terms: D055910:Antibodies, Immobilized / Q000276:immunology; D015374:Biosensing Techniques / Q000295:instrumentation*; D001943:Breast Neoplasms / Q000097:blood* / Q000473:pathology; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000295:instrumentation*; D005898:Glass / Q000737:chemistry; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation*; D007633:Keratins / Q000097:blood / Q000378:metabolism*; D009360:Neoplastic Cells, Circulating / Q000378:metabolism* / Q000473:pathology*
chemical_list: D055910:Antibodies, Immobilized; D007633:Keratins