title: Capture of Tumor Cells on Anti-EpCAM-Functionalized Poly(acrylic acid)-Coated Surfaces.
abstract: The presence of tumor cells in blood is predictive of short survival in several cancers and their isolation and characterization can guide toward the use of more effective treatments. These circulating tumor cells (CTC) are, however, extremely rare and require a technology that is sufficiently sensitive and specific to identify CTC against a background of billions of blood cells. Immuno-capture of cells expressing the epithelial cell adhesion molecule (EpCAM) are frequently used to enrich CTC from blood. The choice of bio conjugation strategy and antibody clone is crucial for adequate cell capture but is poorly understood. In this study, we determined the binding affinity constants and epitope binding of the EpCAM antibodies VU1D-9, HO-3, EpAb3-5, and MJ-37 by surface plasmon resonance imaging (SPRi). Glass surfaces were coated using a poly(acrylic acid) based coating and functionalized with anti-EpCAM antibodies. Binding of cells from the breast carcinoma cell line (SKBR-3) to the functionalized surfaces were compared. Although EpAb3-5 displayed the highest binding affinity HO-3 captured the highest amount of cells. Hence we report differences in the performance of the different antibodies and more importantly that the choice of antibody to capture CTC should be based on multiple assays.
authors.affiliation: Medical Cell Biophysics Group, MIRA Institute for Biomedical Engineering and Technical Medicine, Faculty of Science and Technology, University of Twente , 7522 NB Enschede, The Netherlands.
authors.forename: Kiki C
authors.initials: KC
authors.lastname: Andree
authors.affiliation: Medical Cell Biophysics Group, MIRA Institute for Biomedical Engineering and Technical Medicine, Faculty of Science and Technology, University of Twente , 7522 NB Enschede, The Netherlands.
authors.forename: Ana M C
authors.initials: AM
authors.lastname: Barradas
authors.affiliation: Aquamarijn Micro Filtration BV , IJsselkade 7, 7201 HB Zutphen, The Netherlands.
authors.forename: Ai T
authors.initials: AT
authors.lastname: Nguyen
authors.affiliation: Medical Cell Biophysics Group, MIRA Institute for Biomedical Engineering and Technical Medicine, Faculty of Science and Technology, University of Twente , 7522 NB Enschede, The Netherlands.
authors.forename: Anouk
authors.initials: A
authors.lastname: Mentink
authors.affiliation: Medical Cell Biophysics Group, MIRA Institute for Biomedical Engineering and Technical Medicine, Faculty of Science and Technology, University of Twente , 7522 NB Enschede, The Netherlands.
authors.forename: Ivan
authors.initials: I
authors.lastname: Stojanovic
authors.affiliation: Aquamarijn Micro Filtration BV , IJsselkade 7, 7201 HB Zutphen, The Netherlands.
authors.forename: Jacob
authors.initials: J
authors.lastname: Baggerman
authors.affiliation: Medical Cell Biophysics Group, MIRA Institute for Biomedical Engineering and Technical Medicine, Faculty of Science and Technology, University of Twente , 7522 NB Enschede, The Netherlands.
authors.forename: Joost
authors.initials: J
authors.lastname: van Dalum
authors.affiliation: Laboratory of Organic Chemistry, Wageningen University , Dreijenplein 8, 6703 HB Wageningen, The Netherlands.
authors.forename: Cees J M
authors.initials: CJ
authors.lastname: van Rijn
authors.affiliation: Medical Cell Biophysics Group, MIRA Institute for Biomedical Engineering and Technical Medicine, Faculty of Science and Technology, University of Twente , 7522 NB Enschede, The Netherlands.
authors.forename: Leon W M M
authors.initials: LW
authors.lastname: Terstappen
mesh_terms: D000180:Acrylic Resins / Q000737:chemistry; D000906:Antibodies / Q000378:metabolism*; D000951:Antigens, Neoplasm / Q000378:metabolism; D001943:Breast Neoplasms / Q000175:diagnosis; D045744:Cell Line, Tumor; D019937:Diagnostic Techniques and Procedures / Q000592:standards; D000071858:Epithelial Cell Adhesion Molecule; D006801:Humans; D009360:Neoplastic Cells, Circulating / Q000378:metabolism*
chemical_list: D000180:Acrylic Resins; D000906:Antibodies; D000951:Antigens, Neoplasm; D000071858:Epithelial Cell Adhesion Molecule; C006903:carbopol 940
keywords: antibodies; circulating tumor cells; epithelial cell adhesion molecule (EpCAM); flow model; polycarboxylate coating; surface plasmon resonance