title: Rapid formation of cell-particle complexes via dielectrophoretic manipulation for the detection of surface antigens.
abstract: A rapid and simple method for the fabrication of the island patterns with particles and cells was applied to detect the presence of specific antigens on the cell surface. An upper interdigitated microband array (IDA) electrode was mounted on a lower substrate with the same design to fabricate a microfluidic-channel device for dielectrophoretic manipulation. The electrode grid structure was fabricated by rotating the upper template IDA by 90° relative to the lower IDA. A suspension of anti-CD33 modified particles and HL-60 cells was introduced into the channel. An AC electrical signal (typically 20 V peak-to-peak, 100 kHz) was then applied to the bands of the upper and lower IDAs, resulting in the formation of island patterns at the intersections with low electric fields. Immunoreactions between the antibodies immobilized on the accumulated particles and the CD33 present on the surface of the cells led to the formation of complexes comprising corresponding antigen-antibody pairs. Non-specific pairs accumulated at the intersection, which did not form complexes, were then dispersed after removal of the applied field. The time required for the detection of the formation/dispersion of the complexes is as short as 6 min in the present procedure. Furthermore, this novel cell binding assay does not require pretreatment such as target labeling or washing of the unbound cells.
authors.affiliation: Graduate School of Material Science, University of Hyogo,3-2-1 Kouto, Kamigori, Ako, Hyogo 678-1297, Japan.
authors.forename: Takuma
authors.initials: T
authors.lastname: Horii
authors.affiliation: Graduate School of Material Science, University of Hyogo,3-2-1 Kouto, Kamigori, Ako, Hyogo 678-1297, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Yamamoto
authors.affiliation: Graduate School of Material Science, University of Hyogo,3-2-1 Kouto, Kamigori, Ako, Hyogo 678-1297, Japan; JST-CREST, 5, Sanbancho, Chiyoda, Tokyo 102-0075, Japan. Electronic address: yasu@sci.u-hyogo.ac.jp.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Yasukawa
authors.affiliation: Graduate School of Material Science, University of Hyogo,3-2-1 Kouto, Kamigori, Ako, Hyogo 678-1297, Japan. Electronic address: mizutani@sci.u-hyogo.ac.jp.
authors.forename: Fumio
authors.initials: F
authors.lastname: Mizutani
mesh_terms: D055910:Antibodies, Immobilized / Q000737:chemistry; D015374:Biosensing Techniques / Q000191:economics / Q000295:instrumentation*; D004566:Electrodes; D004586:Electrophoresis / Q000295:instrumentation; D004867:Equipment Design; D018922:HL-60 Cells; D006801:Humans; D007118:Immunoassay / Q000191:economics / Q000295:instrumentation; D046210:Microfluidic Analytical Techniques / Q000191:economics / Q000295:instrumentation*; D063268:Sialic Acid Binding Ig-like Lectin 3 / Q000032:analysis*
chemical_list: D055910:Antibodies, Immobilized; D063268:Sialic Acid Binding Ig-like Lectin 3
keywords: Cell surface antigen; Island pattern; Negative-dielectrophoresis; Particle-cell complex