title: A lipobead microarray assembled by particle entrapment in a microfluidic obstacle course and used for the display of cell membrane receptors.
abstract: Platforms which can display cell membrane ligands and receptors as a microarray library of probes for screening against a target are essential tools in drug discovery, biomarker identification, and pathogen detection. Membrane receptors and ligands require their native bilayer environment to retain their selectivity and binding affinity, and this complicates displaying them in a microarray platform. In this study, a design is developed in which the probes are first incorporated in supported lipid bilayers formed around micron-sized particles (lipobeads), and the microbeads themselves are then arrayed on a surface by hydrodynamic capture in a microfluidic obstacle course of traps. The traps are "V" shaped open enclosures, which are arranged in a wide channel of a microfluidic device, and capture the lipobeads (slightly smaller than the channel height) as they are streamed through the course. Screening assays are undertaken directly in the device after assembly, by streaming a fluorescently labeled target through the device and detecting the bead fluorescence. Conditions are first established for which the supported bilayers on the bead surface remain intact during the capture and assay steps, using fluorescent tags in the bilayer to infer bilayer integrity. Numerical calculations of the hydrodynamic drag coefficient on the entrapped beads are presented in conjunction with the stability experiments to develop criteria for the bilayer stability as a function of the screening assay perfusion rate. Simulations of the flow streamlines are also presented to quantify the trapping efficiency of the obstacle course. Screening assays are illustrated, assaying fluorescently labeled NeutrAvidin with biotin, and labeled cholera toxin with its ganglioside binding ligand, GM1. Sequential capturing of sets of lipobeads (one at a time, and with each set bearing a different probe), followed by indexing the bead positions after each set is entrapped, allows for the construction of an indexed array of multiple probes without the need for particle encoding and is illustrated using the NeutrAvidin-biotin pair. Finally, the lipobead platform is used for quantitatively measuring the kinetic rate constants for the binding of a probe (biotin) to a target (NeutrAvidin).
authors.affiliation: Levich Institute and Department of Chemical Engineering, The City College of the City University of New York, New York, New York 10031, USA.
authors.forename: Xiaoxiao
authors.initials: X
authors.lastname: Chen
authors.forename: Shahab
authors.initials: S
authors.lastname: Shojaei-Zadeh
authors.forename: M Lane
authors.initials: ML
authors.lastname: Gilchrist
authors.forename: Charles
authors.initials: C
authors.lastname: Maldarelli
mesh_terms: D001360:Avidin / Q000032:analysis* / Q000378:metabolism; D001710:Biotin / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002772:Cholera Toxin / Q000032:analysis* / Q000378:metabolism; D004867:Equipment Design; D005456:Fluorescent Dyes / Q000032:analysis; D057446:Hydrodynamics; D008051:Lipid Bilayers / Q000378:metabolism; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D011956:Receptors, Cell Surface / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D011956:Receptors, Cell Surface; C034993:ganglioside receptor; C093483:neutravidin; D001360:Avidin; D001710:Biotin; D002772:Cholera Toxin