title: Highly Selective Capture Surfaces on Medical Wires for Fishing Tumor Cells in Whole Blood.
abstract: The detection of circulating tumor cells (CTCs) in the blood of cancer patients is a challenging task. CTCs are, especially at the early stages of cancer development, extremely rare cells hidden in a vast background of regular blood cells. We describe a new strategy for the isolation of CTCs from whole blood. The key component is a medical wire coated with a multilayer assembly that allows highly specific capture of EpCAM (epithelial cell adhesion molecule) positive CTCs from blood. The assembly is generated in a layer-by-layer fashion through photochemically induced C,H insertion reactions and consists of a protective layer, which shields the contacting solution from the metal, a protein resistant layer, which prevents nonspecific interactions with proteins and a layer containing the EpCAM antibodies. In vitro experiments show that these surfaces can capture tumor cells from whole blood with enrichment factors (specifically vs nonspecifically bound cells) of up to about 3000 compared to the number of leucocytes in the blood. The purity of the isolated cells is greater than 90%. After "fishing" them from the blood, the cells, still bound to the wire, can be genetically analyzed. This demonstrates that this strategy might prove useful for next generation sequencing.
authors.affiliation: Department of Microsystems Engineering, University of Freiburg , Georges-Köhler-Allee 103, D-79110 Freiburg, Germany.
authors.forename: Frank D
authors.identifier: 0000-0002-9126-4196
authors.initials: FD
authors.lastname: Scherag
authors.affiliation: GILUPI GmbH , Hermannswerder 20a, 14473 Potsdam, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Niestroj-Pahl
authors.affiliation: GILUPI GmbH , Hermannswerder 20a, 14473 Potsdam, Germany.
authors.forename: Solveigh
authors.initials: S
authors.lastname: Krusekopf
authors.affiliation: GILUPI GmbH , Hermannswerder 20a, 14473 Potsdam, Germany.
authors.forename: Klaus
authors.initials: K
authors.lastname: Lücke
authors.affiliation: Department of Microsystems Engineering, University of Freiburg , Georges-Köhler-Allee 103, D-79110 Freiburg, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Brandstetter
authors.affiliation: Department of Microsystems Engineering, University of Freiburg , Georges-Köhler-Allee 103, D-79110 Freiburg, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Rühe
mesh_terms: D000906:Antibodies / Q000276:immunology; D055910:Antibodies, Immobilized / Q000276:immunology; D001943:Breast Neoplasms / Q000097:blood / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival; D020099:Coated Materials, Biocompatible; D004797:Enzyme-Linked Immunosorbent Assay; D000071858:Epithelial Cell Adhesion Molecule / Q000276:immunology / Q000378:metabolism; D005260:Female; D006801:Humans; D007938:Leukemia / Q000097:blood / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D009369:Neoplasms / Q000097:blood* / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D009360:Neoplastic Cells, Circulating; D009696:Nucleic Acids / Q000032:analysis; D015203:Reproducibility of Results; D013499:Surface Properties
chemical_list: D000906:Antibodies; D055910:Antibodies, Immobilized; D020099:Coated Materials, Biocompatible; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; D009696:Nucleic Acids