Cell membrane array fabrication and assay technology.
BACKGROUND
Microarray technology has been used extensively over the past 10 years for assessing gene expression, and has facilitated precise genetic profiling of everything from tumors to small molecule drugs. By contrast, arraying cell membranes in a manner which preserves their ability to mediate biochemical processes has been considerably more difficult.


RESULTS
In this article, we describe a novel technology for generating cell membrane microarrays for performing high throughput biology. Our robotically-arrayed supported membranes are physiologically fluid, a critical property which differentiates this technology from other previous membrane systems and makes it useful for studying cellular processes on an industrialized scale. Membrane array elements consist of a solid substrate, above which resides a fluid supported lipid bilayer containing biologically-active molecules of interest. Incorporation of transmembrane proteins into the arrayed membranes enables the study of ligand/receptor binding, as well as interactions with live intact cells. The fluidity of these molecules in the planar lipid bilayer facilitates dimerization and other higher order interactions necessary for biological signaling events. In order to demonstrate the utility of our fluid membrane array technology to ligand/receptor studies, we investigated the multivalent binding of the cholera toxin B-subunit (CTB) to the membrane ganglioside GM1. We have also displayed a number of bona fide drug targets, including bacterial endotoxin (also referred to as lipopolysaccharide (LPS)) and membrane proteins important in T cell activation.


CONCLUSIONS
We have demonstrated the applicability of our fluid cell membrane array technology to both academic research applications and industrial drug discovery. Our technology facilitates the study of ligand/receptor interactions and cell-cell signaling, providing rich qualitative and quantitative information.
Yamazaki
Victoria
V
Synamem Corporation, 863 Mitten Road, Suite 101, Burlingame, CA 94010, USA. vyamazaki@synamem.com
Sirenko
Oksana
O
Schafer
Robert J
RJ
Nguyen
Luat
L
Gutsmann
Thomas
T
Brade
Lore
L
Groves
Jay T
JT
D000818:Animals; D001665:Binding Sites; D001709:Biotechnology / Q000379:methods*; D002454:Cell Differentiation; D002462:Cell Membrane / Q000378:metabolism*; D002772:Cholera Toxin / Q000737:chemistry; D019281:Dimerization; D015195:Drug Design; D004345:Drug Industry; D005677:G(M1) Ganglioside / Q000737:chemistry; D000949:Histocompatibility Antigens Class II / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D008024:Ligands; D008051:Lipid Bilayers / Q000737:chemistry; D008070:Lipopolysaccharides / Q000378:metabolism; D008566:Membranes / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods*; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D012371:Robotics; D019779:Salmonella enterica / Q000378:metabolism; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000378:metabolism; D013678:Technology, Pharmaceutical
D000949:Histocompatibility Antigens Class II; D008024:Ligands; D008051:Lipid Bilayers; D008070:Lipopolysaccharides; D018799:Intercellular Adhesion Molecule-1; D005677:G(M1) Ganglioside; D002772:Cholera Toxin