title: Application of an improved magnetic immunosorbent in an Ephesia chip designed for circulating tumor cell capture.
abstract: In this study, we describe a particular step in developing a microfluidic device for capture and detection of circulating tumor cells-specifically the preparation of an immunosorbent for implementation into the separation chip. We highlight some of the most important specifics connected with superparamegnetic microspheres for microfluidic purposes. Factors such as nonspecific adsorption on microfluidic channels, interactions with model cell lines, and tendency to aggregation were investigated. Poly(glycidyl methacrylate) microspheres with carboxyl groups were employed for this purpose. To address the aforementioned challenges, the microspheres were coated with hydrazide-PEG-hydrazide, and subsequently anti-epithelial cell adhesion molecule (EpCAM) antibody was immobilized. The prepared anti-EpCAM immunosorbent was pretested using model cell lines with differing EpCAM density (MCF7, SKBR3, A549, and Raji) in a batchwise arrangement. Finally, the entire system was implemented and studied in an Ephesia chip and an evaluation was performed by the MCF7 cell line.
authors.affiliation: Department of Biological and Biochemical Sciences, Faculty of Chemical Technology, University of Pardubice, Pardubice, Czech Republic.
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Svobodova
authors.forename: Jana
authors.initials: J
authors.lastname: Kucerova
authors.forename: Julien
authors.initials: J
authors.lastname: Autebert
authors.forename: Daniel
authors.initials: D
authors.lastname: Horak
authors.forename: Lenka
authors.initials: L
authors.lastname: Bruckova
authors.forename: Jean-Louis
authors.initials: JL
authors.lastname: Viovy
authors.forename: Zuzana
authors.initials: Z
authors.lastname: Bilkova
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism; D000951:Antigens, Neoplasm / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000378:metabolism; D000071858:Epithelial Cell Adhesion Molecule; D006801:Humans; D018189:Immunomagnetic Separation / Q000295:instrumentation / Q000379:methods*; D059346:Magnets; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D008863:Microspheres; D009360:Neoplastic Cells, Circulating; D011109:Polymethacrylic Acids / Q000737:chemistry
chemical_list: D055910:Antibodies, Immobilized; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; D000071858:Epithelial Cell Adhesion Molecule; D011109:Polymethacrylic Acids; C042535:polyglycidyl methacrylate
keywords: Biofunctionalization; Circulating tumor cells; EpCAM; Immunomagnetic separation; Poly(glycidyl methacrylate)